• Title/Summary/Keyword: lytic activity

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The Production and Enzymatic Properties of Extracellular Chitinase from Pseudomonas stutzeri YPL-1, as a Biocontrol Agent

  • Lim, Ho-Seong;Kim, Sang-Dal
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.134-140
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    • 1994
  • An antagonistic bacterium Pseudomonas stutzeri YPL-1 liberated extracellular chitinase and $\beta$-1,3-glucanase which are key enzymes in the decomposition of fungal hyphal walls. The lytic enzymes caused abnormal swelling and retreating at the hyphal tips of plant pathogenic fungus Fusarium solani in a dual culture. Scanning electron microscopy revealed the hyphal degradation of F. solani in the regions interacting with P. stutzeri YPL-1. The production of chitinase and properties of a crude preparation of the enzyme from P. stutzeri YPL-1 were investigated. Peak of the chitinase activity was detected after 4 hr of cultivation. The enzyme had optimum temperature and pH of 50$^{\circ}C$ and pH 5.3, respectively. The enzyme was stable in the pH range of 3.5 to 6.0 up to 50$^{\circ}C$. The enzyme was significantly inhibited by metal compounds such as $HgCl_2$, but was stimulated by $CoCl_2$. P. stutzeri YPL-1 produced high levels of the enzyme after 84 hr of incubation. Among the tested carbon sources, chitin was the most effective for the enzyme production, at the concentration level of 3%. As a source of nitrogen, peptone was the best for the enzyme production, at the concentration level of 4%. The maximum amount of enzyme was produced by cultivating the bacterium at a medium of initial pH 6.8.

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The Characteristics of Bacteriophage-resistant Lactococcus lactis subsp. cremoris ATCC 11602-A1 (Lactococcus lactis subsp. cremoris ATCC 11602의 Bacteriophage 내성균주 A1의 특성에 관한 연구)

  • Lee, Chun-Hwa;Kang, Kuk-Hee;Bae, In-Hyu
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.293-298
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    • 1993
  • The ppage resistance mechanism of Lactococcus lactis subsp. cremoris ATCC 11602-A1 was investigated. When parent and A1 were incubated at 30 and 40$^{\circ}C$, A1 grew well and multiplication of phage(MOI=1)on A1 slightly occurred at 40$^{\circ}C$ in contrast with parent. There was a great difference of proteolytic activity between parent and A1, irrespective of the temperature. As a result of ADS treatment oon culture broth, survival rate of A1 was 27% at the lethal concentration of parent and adsorption rate of phage was increased to 95~97%, which was considered to come from the exposure of phage receptor site masked by an unknown component. These results suggest that acridine orange (AO) treatment leads to the modification of cell wall, conferring resistance to high temperature and lytic phage. No change in plasmid profiles of A1 at 30 and 40$^{\circ}C$ were found, which suggests that plasmid is not relative to temperature-resistance of A1.

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Transformation of Cell Wall-weakened Perilla Seedlings Using Phenolic Compound-treated Agrobacterium Cells and Recombinant Protein Expression (페놀화합물 처리 Agrobacterium 및 세포벽 약화 들깨새싹을 이용한 형질전환과 재조합 단백질 발현)

  • Chung, Il-Kyung;Shin, Dong-Il;Park, Hee-Sung
    • KSBB Journal
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    • v.24 no.6
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    • pp.598-601
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    • 2009
  • Perilla [Perilla frutescens (L.) Britt] seedlings are easy to grow and eaten as the health vegetable sprout. Two day old perilla seedlings since germination were given a mild wounding using cell wall lytic NaOH/SDS solution for infiltration with recombinant Agrobacterium cells treated with phenolic compounds. In the analysis of fluorometric GUS gene expression for the transformed perilla seedlings, GUS enzyme activity was the highest by the combined treatments of 50 mM acetosyringone and 0.5% NaOH solution containing 0.01% SDS implying a synergic effect. This result could be successfully applied for demonstrating hepatitis B virus antigen (HBsAg) protein expression.

Modification of Functionality for Lysozyme (Lysozyme의 기능성 개선)

  • Kim, Hyun-Ku
    • Applied Biological Chemistry
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    • v.37 no.6
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    • pp.456-462
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    • 1994
  • Lysozyme-dextran hybrids were prepared by incubated storage at $60^{\circ}C$ and 80% relative humidity for 16 days. The emulsifying properties of the hybrids were about 14 times higher than those of native lysozyme and were about 3 times higher than those of commercial emulsifiers. Lytic activity of the hybrids remained about 83% that of native lysozyme when mesured against Micrococcus lysodeikticus as a substrate. The excellent emulsifying properties of the hybrids were maintained even at pH 3 and were further improved at pH 10. The emulsifying properties of the hybrids were greatly improved by preheating the hybrids at $100^{\circ}C$. In addition the lysozyme-dextran hybrids showed an antimicrobial effect on Gram-negative bacteria.

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The Characteristics, Detection and Control of Bacteriophage in Fermented Dairy Products (발효유제품에서 박테리오파지의 특성, 검출과 제어)

  • Ahn, Sung-Il;Azzouny, Rehab A.;Huyen, Tran Thi Thanh;Kwak, Hae-Soo
    • Food Science of Animal Resources
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    • v.29 no.1
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    • pp.1-14
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    • 2009
  • This study was to review the classification, detection and control of bacteriophage in fermented dairy products. Bacteriophage has lytic and/or lysogenic life cycles. Epidemiologically speaking, detected major phages are c2, 936 and p335. Among them p335 has been the largest concern in dairy industry. Traditionally, various analytical technologies, such as spot, starter activity, indicator test, ATP measurement and conductimetric analysis, have been used for the phage detection. In recent years, advanced methods such as flow cytometric method, petrifilm, enzyme linked immunosorbent assay (ELISA) and multiflex PCR diagnostic kit have been deveoloped. The phage contamination has been controlled by using heat, high-pressure treatment, and the combinations of heat and pressure, and/or chemical. Also some starter cultures with phage-resistant character have been developed to minimize the concentration of phages in dairy product. Bacteriophage inhibition media such as calcium medium was also mentioned. To prevent the contamination of bacteriophage in dairy industry, further researches on the detection and control of phage, and phage resistant starters are necessary in the future.

Antagonistic effects Na+ and Mg2+ on the structure, function, and stability of mycobacteriophage L1 repressor

  • Bandhu, Amitava;Ganguly, Tridib;Chanda, Palas K.;Das, Malabika;Jana, Biswanath;Chakrabarti, Gopal;Sau, Subrata
    • BMB Reports
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    • v.42 no.5
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    • pp.293-298
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    • 2009
  • Temperate mycobacteriophage L1 encodes an unusual repressor (CI) for regulating its lytic-lysogenic switching and, in contrast to the repressors of most temperate phages, it binds to multiple asymmetric operator DNAs. Here, ions like $Na^+$, $Cl^-$, and $acetate^-$ ions were demonstrated to facilitate the optimal binding of CI to cognate operator DNA, whereas $K^+$, $Li^+$, ${NH_4}^+$, $Mg^{2+}$, $carbonate^{2-}$, and $citrate^{3-}$ ions significantly affected its operator binding activity. Of these ions, $Mg^{2+}$ unfolded CI most severely at room temperature and, compared to $Mg^{2+}$, $Na^+$ provided improved thermal stability to CI. Furthermore, the intrinsic tryptophan fluorescence of CI was changed notably upon replacing $Na^+$ with $Mg^{2+}$ and these opposing effects of $Mg^{2+}$ and $Na^+$ were also noticed in their actions on the C-terminal fragment (CTD) of CI. Taken together, $Na^+$ appeared to be more appropriate than $Mg^{2+}$ for maintaining the biologically active conformation of CI needed for its optimal binding to operator DNA.

Notch Signal Transduction Induces a Novel Profile of Kaposi's Sarcoma-Associated Herpesvirus Gene Expression

  • Chang Hee-Soon
    • Journal of Microbiology
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    • v.44 no.2
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    • pp.217-225
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    • 2006
  • Kaposi's sarcoma-associated herpesvirus (KSHV) RTA transcription factor is recruited to its responsive elements through interaction with RBP-Jk that is a downstream transcription factor of the Notch signaling pathway that is important in development and cell fate determination. This suggests that KSHV RTA mimics cellular Notch signal transduction to activate viral lytic gene expression. Here, I demonstrated that unlike other B lymphoma cells, KSHV -infected primary effusion lymphoma BCBL1 cells displayed the constitutive activation of ligand-mediated Notch signal transduction, evidenced by the Jagged ligand expression and the complete proteolytic process of Notch receptor I. In order to investigate the effect of Notch signal transduction on KSHV gene expression, human Notch intracellular (hNIC) domain that constitutively activates RBP-Jk transcription factor activity was expressed in BCBL1 cells, TRExBCBL1-hNIC, in a tetracycline inducible manner. Gene expression profiling showed that like RTA, hNIC robustly induced expression of a number of viral genes including KS immune modulatory gene resulting in downregulation of MHC I and CD54 surface expression. Finally, the genetic analysis of KSHV genome demonstrated that the hNIC-mediated expression of KS during viral latency consequently conferred the downregulation of MHC I and CD54 surface expression. These results indicate that cellular. Notch signal transduction provides a novel expression profiling of KSHV immune deregulatory gene that consequently confers the escape of host immune surveillance during viral latency.

A Revised Assay for Monitoring Autophagic Flux in Arabidopsis thaliana Reveals Involvement of AUTOPHAGY-RELATED9 in Autophagy

  • Shin, Kwang Deok;Lee, Han Nim;Chung, Taijoon
    • Molecules and Cells
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    • v.37 no.5
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    • pp.399-405
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    • 2014
  • Autophagy targets cytoplasmic cargo to a lytic compartment for degradation. Autophagy-related (Atg) proteins, including the transmembrane protein Atg9, are involved in different steps of autophagy in yeast and mammalian cells. Functional classification of core Atg proteins in plants has not been clearly confirmed, partly because of the limited availability of reliable assays for monitoring autophagic flux. By using proUBQ10-GFP-ATG8a as an autophagic marker, we showed that autophagic flux is reduced but not completely compromised in Arabidopsis thaliana atg9 mutants. In contrast, we confirmed full inhibition of auto-phagic flux in atg7 and that the difference in autophagy was consistent with the differences in mutant phenotypes such as hypersensitivity to nutrient stress and selective autophagy. Autophagic flux is also reduced by an inhibitor of phosphatidylinositol kinase. Our data indicated that atg9 is phenotypically distinct from atg7 and atg2 in Arabidopsis, and we proposed that ATG9 and phosphatidylinositol kinase activity contribute to efficient autophagy in Arabidopsis.

Effects of Juglans regia Complex Extract on Osteoclast Differentiation from Bone Marrow Derived Macrophage (호두복합추출물이 골수유래대식세포의 파골세포 분화에 미치는 효과)

  • Kong, Hae Jin;Kang, Jae Hui;Ryu, Hwa Yeon;Lee, Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.33 no.3
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    • pp.169-174
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    • 2019
  • The purpose of this study was to evaluate the inhibitory effects of Juglans regia complex extract(JCE) consisted of Juglans regia, Eucommia ulmoides, Eleutherococcus senticosus and Zingiber officinale on osteoclast differentiation. Cell toxicity test by using CCK-8, TRAP activity and TRAP positive multi-nucleated cell counting were performed to evaluate inhibitory effect on differentiation of osteoclast from bone marrow derived macrophages(BMMs) induced by receptor activator of nuclear $factor-{\kappa}B$ ligand(RANKL). As a result, JCE inhibited RANKL-induced osteoclast differentiation in BMMs dose-dependently without cytotoxicity. These results suggest that JCE may have a potential role for treating bone lytic diseases such as osteoporosis.

Cellular responses and proteomic analysis of hemolytic Bacillus cereus MH-2 exposed to epigallocatechin gallate (EGCG) (Epigallocatechin Gallate (EGCG)에 노출된 용혈성 Bacillus cereus MH-2의 세포 반응 및 프로테옴 분석)

  • Kim, Dong-Min;Park, Sang-Kook;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.260-268
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    • 2016
  • The aim of this work was to investigate the cellular responses and proteomic analysis of Bacillus cereus MH-2 exposed to EGCG. Strain MH-2 was isolated from commercial Ssamjang and has the hemolytic activity. Survival of the MH-2 strain with time in the presence of different concentrations of EGCG under sublethal conditions was monitored. The amount of alginate from MH-2 strain decreased depending on the increasing concentrations of EGCG and increased depending on the exposure time at any particular EGCG concentration. Analysis of SDS-PAGE and Western blot using anti-DnaK and anti-GroEL revealed that two stress shock proteins, 70 kDa DnaK and 60 kDa GroEL were found to decrease in proportion to the EGCG concentration in exponentially growing cultures. Scanning electron microscopic analysis demonstrated the presence of protrusions and fused rod forms on the cells treated with EGCG. 2-DE of soluble protein fractions from MH-2 cultures showed 20 protein spots changed by EGCG exposure. These proteins involved in enterotoxins (hemolysin BL lytic component L1 and hemolysin BL-binding protein), chaperons (DnaK and GroEL), cell defense (peptidase M4 family proteins), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. These results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on B. cereus MH-2.