• Title/Summary/Keyword: bacterial enzymes

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Current Status on Molecular Genetic Study and Comparative Genomic Analysis of Virulence Related Genes in Xanthomonas oryzae pv. oryzae (벼 흰잎마름병균(Xanthomonas oryzae pv. oryzae)의 병원성 유전자의 분자유전학적 연구현황 및 비교유전체 분석)

  • Kang, Hee-Wan;Park, Young-Jin;Lee, Byeong-Moo
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.1-9
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    • 2008
  • Xanthomonas oryzae pv. oryzae (Xoo) is the causal agent of bacterial blight on rice. In this paper, current status on molecular genetic study of major virulence genes, hypersensitive response and pathogenicity (hrp), productions of extracellular polysaccharide (EPS), extracellular enzymes and lipopolysaccharides (LPS), avr genes were reviewed. The IS elements with 611 copies including 133 ORF IS were inserted in various regions of the Xoo genome and in expecially regions franking virulence genes. Whole genome sequence of X. oryzae pv. oryzae KACC10331 were used for defining genetic organization of the virulence genes. Futhermore, the virulence genes in Xoo genome were compared to those of other Xanthomonas species in Blast GenBank data base.

Molecular Diversity of Rhizobacteria in Ginseng Soil and Their Plant Benefiting Attributes (인삼토양 내 근권세균의 다양성 및 식물에의 유용 특성)

  • Hong, Eun Hye;Lee, Sun Hee;Vendan, Regupathy Thamizh;Rhee, Young Ha
    • Korean Journal of Microbiology
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    • v.48 no.4
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    • pp.246-253
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    • 2012
  • The purpose of this study was to investigate the molecular diversity of rhizobacteria associated with ginseng of varying age levels and their plant benefiting attributes. A total of 143 different isolates belonging to 15 different bacterial genera were recovered. Although variation was found in the rhizobacterial community due to age of the plant, majority of bacteria belong to Firmicutes (58%). In which, Bacillus was found to be the predominant genus irrespective of age of the ginseng. To assess the plant benefiting attributes, 30 representative isolates were selected. The results indicated that some of the isolates could exhibit multiple plant growth promoting traits like secretion of cell wall degrading enzymes, production of indole-3-acetic acid, synthesis of siderophores, solubilization of phosphates and soil pathogens inhibition. It can be suggested that strains of B. subtilis, B. amyloliquefaciens, B. velezensis, and B. licheniformis were positive for all the above traits, which have potential to be used as plant growth promoting inoculants to improve ginseng crop in the future.

Isolation of an Acinetobacter junii SY-01 Strain Producing an Extracellular Lipase Enantioselectively Hydrolyzing Itraconazole Precursor, and Some Properties of the Lipase

  • Yoon, Moon-Young;Shin, Pyong-Kyun;Han, Ye-Sun;Lee, So-Ha;Park, Jung-Keug;Cheong, Chan-Seong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.97-104
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    • 2004
  • Water-sludge bacteria were screened to find a lipase enantioselectively hydrolyzing itraconazole precursor, which is well known as the starting material of antifungal drug agents. A bacterial strain was isolated and identified as Acinetobacter junii SY-01. After the strain was cultivated, the enzyme was purified 39.4-fold using ultrafiltration and gel filtration through a Sephadex G-100 chromatographic column and the activity yield was 34.9%. The molecular weight of the enzyme was about 40 kDa, as measured by SDS-PAGE, and the optimum pH was 7.0- 9.0 and stable at pH 6.0- 9.0. The optimum temperature was 45- $5^{\circ}C$, and 73% of the enzymes activity remained after incubation at 70% for 1 h. Enzyme activity was enhanced by gall powder, sodium deoxycholate, a cationic detergent Tween 80, and a non-ionic detergent Triton X-100, but was markedly inhibited by metal ions such as $Hg^{2+},Cu^{2+},Ni^{2+}/,Ca^{2+}$, and an anionic-surfactant sodium dodecylsulfate. The $K_{m}$ values for (R)- and (S)-enantiomers of the itraconazole precursor were 0.385 and 21.83 mM, respectively, and the $V_{max} values ($\mu$Mㆍmin^{-1}.)$ were 6.73 and 6.49, respectively. The acetyl group among the different acyl moieties of itraconazole precursor showed the highest enantioselectivity for the hydrolysis by the Acinetobacter junii SY-01 lipase, and the lipase from Acinetobacter junii SY-01 displayed better enantioselectivity than that of commercially available lipases and esterases.

Halotolerant Spore-Forming Gram-Positive Bacterial Diversity Associated with Blutaparon portulacoides (St. Hill.) Mears, a Pioneer Species in Brazilian Coastal Dunes

  • Barbosa Deyvison Clacino;Irene Von Der Weid;Vaisman Natalie;Seldin Lucy
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.193-199
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    • 2006
  • Halotolerant spore-forming Gram-positive bacteria were isolated from the root, rhizosphere, and non-rhizosphere soil of Blutaparon portulacoides. The different isolates were characterized genetically using an amplified ribosomal DNA restriction analysis (ARDRA), and phenotypically based on their colonial morphology, physiology, and nutritional requirements. Three different 16S rRNA gene-based genotypes were observed at a 100% similarity using the enzymes HinfI, MspI, and RsaI, and the phenotypic results also followed the ARDRA groupings. Selected strains, representing the different ARDRA groups, were analyzed by 16S rDNA sequencing, and members of the genera Halobaeillus, Virgibacillus, and Oceanobacillus were found. Two isolates showed low 16S rDNA sequence similarities with the closest related species of Halobacillus, indicating the presence of new species among the isolates. The majority of the strains isolated in this study seemed to belong to the species O. iheyensis and were compared using an AP-PCR to determine whether they had a clonal origin or not. Different patterns allowed the grouping of the strains according to Pearson's coefficient, and the resulting dendrogram revealed the formation of two main clusters, denoted as A and B. All the strains isolated from the soil were grouped into cluster A, whereas cluster B was exclusively composed of the strains associated with the B. portulacoides roots. This is the first report on the isolation and characterization of halotolerant spore-forming Gram-positive bacteria that coexist with B. portulacoides. As such, these new strains may be a potential source for the discovery of bioactive compounds with industrial value.

Overexpression, Crystallization, and Preliminary X-Ray Crystallographic Analysis of the Alanine Racemase from Enterococcus faecalis v583

  • Priyadarshi, Amit;Lee, Eun-Hye;Sung, Min-Woo;Kim, Jae-Hee;Ku, Min-Je;Kim, Eunice Eun-Kyeong;Hwang, Kwang-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.55-58
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    • 2008
  • Alanine racemase, a bacterial enzyme belonging to the fold-type III group of pyridoxal 5'-phosphate (PLP)-dependent enzymes, has been shown to catalyze the interconversion between L- and D-alanine. The alanine racemase from the pathogenic bacterium Enterococcus faecalis v583 has been overexpressed in E. coli and was shown to crystallize an enzyme at 295 K, using polyethylene glycol (PEG) 8000 as a precipitant. X-ray diffraction data to $2.5{\AA}$ has been collected using synchrotron radiation. The crystal is a member of the orthorhombic space group, $C222_1$ with unit cell parameter of a=94.634, b=156.516, $c=147.878{\AA},\;and\;{\alpha}={\beta}={\gamma}=90{\AA}$. Two or three monomers are likely to be present in the asymmetric unit, with a corresponding $V_m\; of\;3.38{\AA}^3\;Da^{-1}\;and\;2.26{\AA}^3\;Da^{-1}$ and a solvent content of 63.7% and 45.5%, respectively.

Role of Silver-containing Carboxymethyl Cellulose Dressing in the Management of Exudative Infected Wounds (삼출액과 감염이 동반된 창상의 치료에 있어 은 함유 카르복시메틸 셀룰로즈 드레싱의 역할)

  • Lee, Ji Hyuck;Lee, Eun Sang;Kang, So Ra
    • Archives of Plastic Surgery
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    • v.34 no.3
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    • pp.305-313
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    • 2007
  • Purpose: Wound healing is a result of complex processes whose components, such as cells, extracellular matrix, proteolytic enzymes, and their inhibitors receive effects from immune compartments, cytokines, chemokines, and growth factors. Impairment of normal physiologic response to wounding makes nonhealing chronic wounds. Wound infection and exacerbated proteolytic process may induce uncontrolled tissue degradation or exudates formation, which may result in the development of a nonhealing chronic wound. Thus proper management of wound infection and exudates is critical to prevent and treat nonhealing wound. The aim of this study is to evaluate effects of Aquacel AG, silver-containing carboxymethylcellulose dressing on treatment for exudative infected wound. Methods: The study included 31 patients with nonhealing wound. Wound was dressed with Aquacel AG. The effect of dressing was investigated by serial bacterial culture and wound exudates assessment. Each infection and exudates control time was determined and statistically analyzed. Results: Wound infection and exudates were effectively managed using Aquacel AG dressing. Mean infection and exudates control time were $3.4{\pm}1.2$ and $5.7{\pm}1.4$ weeks, respectively. Statistical analysis of the data indicated that infection control time correlated positively to age and exudates control time (p<0.05). Conclusion: There is as yet no ideal dressing for the topical treatment of chronic nonhealing wound. But silver-containing carboxymethylcellulose dressing can be used effectively for exudative, infected nonhealing wound.

Performance Enhancement of Anaerobic Treatment of Waste Sludge by Chemical Pretreatment (화학적 전처리를 통한 혐기성 슬러지 처리효율의 향상)

  • 허준무;박종안;손부순
    • Journal of environmental and Sanitary engineering
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    • v.13 no.1
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    • pp.16-25
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    • 1998
  • Laboratory-scale experiment using anaerobic fluidized bed reactor was carried out to investigate the prehydrolysis step with caustic soda on the treatment efficiency of anaerobic sludge treatment, since the overall rate-limiting step for the complete anaerobic digestion of sludge was the hydrolysis step by extracellular bacterial enzymes of insoluble polymeric molecules. Reactors received a sludge which had not been pretreated, a 50-50 mixture of pretreated and untreated sludge, and the fully pretreated sludge. Hydraulic retention time of 10, 5, 2.5 days and 1 day were applied with an respective equivalent organic loading rate of 1.17, 2.23, 4.17, 11.24 gCOD/L/d. Reactor with the untreated sludge did not archieve adequate digestion even at the HRT of 5 days, and reactor, which received the 50-50 mixture, operated well at the HRT of 5 days, but began to show signs of unstable digestion at the HRT of 2.5 days. While, reactor, which was fed the hydrolyzed sludge, operated reasonably well at the 2.5 days, but was showing somewhat decrease in removal efficiencies. Results, therefore, have substantiated that the limiting reaction in the anaerobic treatment process is hydrolysis. The soluble COD did not significantly accumulate in the reactor as organic acid form, even when they were highly stressed. It was believed that this resistance to a build-up of organic acids and soluble COD behavior was mainly due to the maintenance of the methane bacteria in the fixed-film system which prevents washout as the organic loading increased. The anaerobic fluidized bed reactor was therefore effective for the digestion of waste activated sludge at short HRT.

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Isolation and Identification of Feather-Degrading Bacteria for Biotechnological Applications of Keratinaceous Protein Waste (케라틴 단백질 폐기물의 생물공학적 적용을 위한 우모 분해세균의 분리 및 동정)

  • 손홍주;김용균;박연규
    • Journal of Life Science
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    • v.14 no.2
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    • pp.229-234
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    • 2004
  • Feathers, which are almost pure keratin protein, are produced in large amounts as a waste by-product at poultry-processing plants. Keratinolytic enzymes may have important uses in biotechnological processes involving keratin-containing wastes from poultry and leather processes. In this study, screening and identification of keratin-degrading bacteria were investigated. Five keratin-degrading bacterial strains (F3-1, F3-4, F7-1, C1-1, C1-2) were isolated from compost and decayed chicken feather. On the basis of morphological, physiological studies, and Biolog system, all isolates were identified as the genus Bacillus. Among them, the strain F7-1 had the highest feather-degrading activity and was selected for further taxonomical study. Phylogenetic analysis of strain F7-1 based on comparison of 165 rDNA sequences revealed that this strain is closely related to Bacillus megaterium.

Scutellarein Reduces Inflammatory Responses by Inhibiting Src Kinase Activity

  • Sung, Nak Yoon;Kim, Mi-Yeon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.5
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    • pp.441-449
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    • 2015
  • Flavonoids are plant pigments that have been demonstrated to exert various pharmacological effects including anti-cancer, anti-diabetic, anti-atherosclerotic, anti-bacterial, and anti-inflammatory activities. However, the molecular mechanisms in terms of exact target proteins of flavonoids are not fully elucidated yet. In this study, we aimed to evaluate the anti-inflammatory mechanism of scutellarein (SCT), a flavonoid isolated from Erigeron breviscapus, Clerodendrum phlomidis and Oroxylum indicum Vent that have been traditionally used to treat various inflammatory diseases in China and Brazil. For this purpose, a nitric oxide (NO) assay, polymerase chain reaction (PCR), nuclear fractionation, immunoblot analysis, a kinase assay, and an overexpression strategy were employed. Scutellarein significantly inhibited NO production in a dose-dependent manner and reduced the mRNA expression levels of inducible NO synthase (iNOS) and tumor necrosis factor (TNF)-${\alpha}$ in lipopolysaccharide (LPS)-activated RAW264.7 cells. In addition, SCT also dampened nuclear factor (NF)-${\kappa}B$-driven expression of a luciferase reporter gene upon transfection of a TIR-domain-containing adapter-inducing interferon-${\beta}$ (TRIF) construct into Human embryonic kidney 293 (HEK 293) cells; similarly, NF-${\kappa}B$ nuclear translocation was inhibited by SCT. Moreover, the phosphorylation levels of various upstream signaling enzymes involved in NF-${\kappa}B$ activation were decreased by SCT treatment in LPS-treated RAW264.7 cells. Finally, SCT strongly inhibited Src kinase activity and also inhibited the autophosphorylation of overexpressed Src. Therefore, our data suggest that SCT can block the inflammatory response by directly inhibiting Src kinase activity linked to NF-${\kappa}B$ activation.

Intraspecific Protoplast Fusion of Brevibacterium and Intergeneric Protoplast Fusion between Brevibacterium flavum and Corynebacterium glutamicum and the Metabolic Control of L-Lysine Biosynthesis in Improved Bacterial Strains (Brevibacterium flavum의 동종간 및 Corynebacterium glutamicum과의 이속간 원형질체 융합 및 개량균주의 L-Lysine 생합성의 대사제어)

  • Park, Chung;Im, Beon-Sam;Jeon, Moon-Jin
    • Microbiology and Biotechnology Letters
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    • v.15 no.2
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    • pp.104-111
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    • 1987
  • As a trial method of breeding L-lysine producing strains, the intraspecific protoplast fusion bet-ween Brevibacterium flavum ATCC 21528R and Brevibacterium flavum ATCC 21529S and the intergeneric protoplast fusion between Brevibacterium flavum ATCC 21528R and Corynebacterium glutamicum ATCC 13058S were performed. The optimum conditions for protoplast formation of these strains were examined and the effect of plasma expander on regeneration and/or fusion was also observed. Both fusants No. CH23 and No. CH4l showed higher productivity of L-lysine than those of parental cells under the optimum cultural conditions at a rate of 21% and 8.9%, respectively. And, activity of several enzymes in L-lysine biosynthetic pathway including aspartokinase, a rate-limiting enzyme, was determined. Besides, metabolic control mechanism of L-lysine biosynthesis in fusant No. CH23 and in No. CH41 was investigated to compare with that of parental strains.

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