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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Microbiology and Biotechnology Letters
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Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 33, Issue 4 - Dec 2005
Volume 33, Issue 3 - Sep 2005
Volume 33, Issue 2 - Jun 2005
Volume 33, Issue 1 - Mar 2005
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Construction of a Lactococcal Shuttle/Expression Vector Containing a
-Galactosidase Gene as a Screening Marker
Han Tae Un ; Jeong Do-Won ; Cho San Ho ; Lee Jong-Hoon ; Chung Dae Kyun ; Lee Hyong Joo ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 241~247
A new lactococcal shuttle/expression vector for lactococci, pWgal13T, was constructed using a
-galactosi-dase gene (lacZ) from Lacfococcus lactis ssp. lactis ATCC 7962 as a screening marker. The pWgal 13T was introduced into Escherichia coli DH5a and L. lactis MG1363, and was easily detected by the formation of blue colonies on a medium containing X-gal without any false transformants. Also, the quantitatively lacZ activity of pWgal13T was measured in L. lactis ssp. cremoris MG1363, and was found to be four times higher than that of L. lactis ssp. lactis ATCC7962 grown on a medium containing glucose, which shows that the lacZ gene of pWgal13T can be used for the efficient screening of L. lactis on general media. The pWgal13T was equipped with a lactococcal replicon of pWV01 from L. lactis Wg2, the new promoter P13C from L. lactis ssp. cremoris LM0230, multiple cloning sites, and a terminator for the expression of a relevant gene. The vee-tor pWgal13T was used for the expression of the EGFP gene in E. coli and L. lactis. These results show that the lactococcal expression/shuttle vector constructed in the present study can be used for the production of foreign proteins in E. coli and L. lactis.
Characterization of Bacteria Isolated from Rotted Onions (Allium cepa)
Lee Chan-Jung ; Lim Si-Kyu ; Kim Byung-Chun ; Park Wan ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 248~254
One hundred thirty nine bacteria were isolated from rotten onions collected from main producing districts, Chang-Nyung, Eui-Ryung, and Ham-Yang in Korea. The
(25 strains) of bacterial isolates have carboxymethylcellulase (CMCase) activity and the
(74 strains) have polygalacturonase (PGase) activity. Thirty one among randomly selected 45 strains of PGase producing bacteria have pathogenicity to onions. The isolates were classified into Pseudomonas sp. (18 strains), Bacillus sp. (11 strains), Yers-inia sp. (7 strains), and others (9 strains) on the basis of FAMEs patterns. Eighteen strains of Pseudomonas sp. were mainly divided into three cluster in the dendrogram and only the two clusters of them showed pathogenicity to onions. CMCase and PGase activities of Pseudomonas sp. weaker than those of Bacillus sp.. However, the pathogenicity of pseudomonas sp. to soften onions was stronger than that of Bacillus sp. Inoculation of
cfu of Pseudomonas sp. gives rise to softening of onions. Pseudomonas sp. was identified as Pseudomonas gladioli by biochemical and physiological characteristics. P. gladioli is the first reported bacterium as a pathogen of onion in Korea. In low temperature, P. gladioli showed better growth and higher PGase activity than those of Bacillus sp. identified as Bacillus subtilis. And pH 9.0 is optimal pH for PGase activity of B. subtilis while that of P. gladioli is pH
which is the acidity of onions. Taken together, P. gladioli may be a main pathogene of onion rot during the cold storage condition.
Interaction between IgE-Dependent Histamine-Releasing Factor and Triosephosphate Isomerase in HeLa Cells
Moon Ji-Ae ; Kim Hwa-Jung ; Lee Kyunglim ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 255~259
IgE-dependent histamine-releasing factor (HRF) is found extracellularly to regulate the degranulation process of histamine in mast cells and basophils and known to play a predominant role in the pathogenesis of chronic allergic disease. HRF has been also identified in the intracellular region of the cell. Previously, we reported that HRF interacts with the 3rd cytoplasmic domain of the alpha subunit of Na,K-ATPase. To understand the molecular mechanism of the regulation of Na, K-ATPase activity by HRF, we investigated the interaction between HRF and TPI since TPI was obtained as HRF-interacting protein in HeLa cDNA library, using yeast two hybrid screening. Domain mapping study of the interaction between HRF and TPI revealed that the C-terminal region of the residue 156-249 of TPI is involved in the interaction with HRF. The interaction between HRF and TPI was confirmed by immunoprecipitation from HeLa cell extracts. Our results suggest that TPI is a HRF-binding protein and the interaction between HRF and TPI nay thus affect Na, K-ATPase activity.
PKC Isotype that Affects the Interaction of HRF with Na, K-ATPase
Sohn Wern-Joo ; Lee Kyunglim ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 260~266
IgE-dependent histamine releasing factor (HRF), previously known as P23/P21 or translationally controlled tumor protein (TCTP), induces the degranulation of histamine in mast cell and basophil. Yeast two hybrid results showed that HRF interacts with the alpha subunit of Na, K-ATPase, suggesting that HRF is a regulator for governing the activity of Na, K-ATPase. In this study, we examined the interaction of HRF and Wa,K-ATPase after treatments of various PKC isotype inhibitors. Membrane fractionation, pull-down assay and immunoprecipitation results showed that PKC
subunits are involved in the phosphorylation of HRF. However, these results did not correlate with the results of histamine release assay since histamine release assay results suggested that some PKC isotype inhibitors induced the histamine release in RBL-2H3 cell.
Expression of Tkermomonoepora fusea Exoglucanase in Saccharomyces cerevisiae and Its Application to Cellulose Hydrolysis
Park Hyun-Soon ; Kim Hyun-Chul ; Shin Dong-Ha ; Kim Joong-Kyun ; Nam Soo-Wan ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 267~273
To develop effective and powerful probiotic, Saccharomyces cerevisiae strains producing cellulolytic enzymes were genetically brooded. For the production of exoglucanase, the plasmid pVT-TExo (8.8 kb) was constructed, in which Thermomonosporafusca exoglucanase gene (E3) was under the control of ADHl promoter, and introduced into S. cerevisiae SEY2102. When the transformant, S. cerevisiae SEY2102/pVT-TExo, was cultivated on YPD medium, the total expression level of avicelase reached about 190 unit/l. The secretion efficiency and plasmid stability were about
, respectively. Recombination exoglucanase enzyme bound to avicel better than Clostridium endoglucanase (CelA) and Trichoderma endoglucanase (C4) enzymes. The mixing ratio of E3 and CelA displaying the best synergistic hydrolysis for avicel was observed at 4:1. The mixture of endoglucanase (CelA) and exoglucanase (E3) resulted in 3.2-fold increase of avicelase activity and 2.5-fold enhanced production of sugar production from avicel, compared to the single enzyme treatment.
Molecular Cloning and Expression of a Gene for Outer Membrane Protein H in Pasteurella multocida (A:3) : Production of Antisera against the OmpH
Kim Younghwan ; Hwang Heon ; Lee Sukchan ; Park Eun-Seok ; Yoo Sun-Dong ; Lee Jeongmin ; Yang Joo-Sung ; Kwon MooSik ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 274~280
Pasteurella multocida is known to cause widespread infections in husbandry. To induce homologous and heterologous immunity against the infections, outer membrane proteins (OMPs) in the envelope of P. multocida are thought to be attractive vaccine candidates. Outer membrane protein H is considered as the major component of OMPs. In this study, a gene for OmpH was isolated from pathogenic P. multocida serogroup A. The gene was composed of 1,047 nucleotides coding 348 amino acids with signal peptide of 20 amino acids. The amino acid composition showed about 80 to 98 per cent sequence homologies among other 10 strains of P. multocida serogroup A, reported so far. A recombinant ompH, from which signal peptide was truncated, was generated using pRSET A to name 'pRSET A/OmpH-F2'. The pRSET A/OmpH-F2 was well expressed in E. coli BL21(DE3). The truncated OmpH was purified using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Its molecular weight was registered to be 40 kDa on SDS-PAGE gel. In order to generate immunesera against the OmpH, 50 ug of the protein was intraperitoneally injected into mice three times. The anti-OmpH immuneserum recognized about
ng quantity of the purified OmpH. It can be used for an effective vaccine production to prevent fowl cholera caused by pathogenic P. multocida (Serogroup A).
Production of Inulooligosaccharides by Endoinulinase Expressed in Saccharomyces cerevisiae
Kim Hyun-Chul ; Kim Hyun-Jin ; Kim Byung-Woo ; Kwon Hyun-Ju ; Nam Soo-Wan ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 281~287
The endoinulinase gene (inu, 2.733 kb, EC 220.127.116.11) from Paenibacillus polymyxa was subcloned into an Escherichia coli-yeast shuttle vector with GALl promoter for the expression in Saccharomyces cerevisiae. The constructed plasmid, pYGENIU27 (8.6 kb) was introduced into S. cerevisiae SEY2102 cell and then the yeast transformant was selected on the synthetic defined media lacking uracil and on the inulin-containing media. The recombinant endoinulinase was predominantly localized in the periplasmic space of the yeast cell. The total activity of the endoinulinase reached 1.81 unit/ml by cultivation of yeast transformant on YPDG medium. The optimized conditions determined for the inulooligosaccharides (IOSs) production from inulin were as follows; pH, 8.0; reaction temperature,
; inulin source, Jerusalem artichoke. Enzyme activity was stably maintained up to the pH of 10.0. Under the optimized condition and with endoinulinase of 36 unit/g-inulin, IOSs started to be produced after 10 min of enzymatic reaction. By the reaction with inulin, IOSs consisting of inulobiose (F2), inulotriose (F3), and inulotetraose (F4) were produced and F3 was the major product. Consequently, these data would be used as a fundamental parameters for the production of functional sweetener IOSs from inulin by recombinant yeast endoinulinase.
Increase of Bioactive Flavonoid Aglycone Extractable from Korean Citrus Peel by Carbohydrate-Hydrol-ysing Enzymes
Ahn Soon-Cheol ; Kim Min-Soo ; Lee Sun-Hi ; Kang Ju-Hyung ; Kim Bo-Hye ; Oh Won-Keun ; Kim Bo-Yeon ; Ahn Jong-Seog ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 288~294
Flavonoid compounds show several biological activities and generally exist in the forms of glycones linking sugar moiety to main structure. Flavonoid glycones such as naringin and hesperidin in korean citrus peel are slower absorbed and consequently less active than their aglycone, naringenin and hesperetin, respectively. Therefare to increase the content of flavonoid aglycone in korean citrus peel, we used commercial carbohydrate-hydrolysing enzymes, AMG 300 L, Pectinex 100 L, and Viscozyme for transforming flavonoid glycones to aglycones. Optimal conditions of enzyme reaction were pH 5.0-7.0,
enzyme, and 24-48 hrs. The content of naringenin and hesperetin as flavonoid aglycones in untreated citrus peel is
of dried citrus peel. In case of enzyme-treated citrus peel the content of naringenin and hesperetin increased to
of dried citrus peel, respectively. Finally the content of flavonoid aglycones could be extracted to 10-80 times. Now enzyme-treated citrus peel may be applied to use for functional food because of its higher flavonoid aglycones as more active compounds.
Isolation and Characterization of Three Kinds of Lipopeptides Produced by Bacillus subtilis JKK238 from Jeot-Kal of Korean Traditional Fermented Fishes
Yoon Sang-Hong ; Kim Jung-Bong ; Lim Yoong-Ho ; Hong Seong-Ryeul ; Song Jae-Kyeung ; Kim Sam-Sun ; Kwon Soon-Wo ; Park In-Cheol ; Kim Soo-Jin ; Yeo Yun-Soo ; Koo Bon-Sung ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 295~301
About seven hundred bacterial strains were collected from Jeot-Kal, a Korean traditional fermented fishes, in various Korean districts. One of the strains designated JKK238 has its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The JKK238 strain was isolated from Oh-Jeot, a kind of fermented shrimps, of Kangkyeung in Korea, and was identified as Bacillus subtilis based on its physiological characteristics, fatty acids compositions of cellular wall, and 16S rDNA sequence analysis. We isolated simply antimicrobial lipopeptides (AMLP) by
ammonium sulfate precipitation of 3 days-old tryptic soy broth cultures of the JKK238 strain. Further analysis of AMLP revealed that B. subtilis JKK238 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families simultaneously. Above results indicate that the JKK238 strain can be added to the limited number B. subtilis strains reported to co-produce the three kinds of lipopeptide families.
Assessment of Inactivation for Campylobacter spp. Attached on Chicken Meat
Jang Keum-Il ; Jeong Heon-Sang ; Kim Chung-Ho ; Kim Kwang-Yup ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 302~307
The inactivation efficiency of Campylobacter jejuni were assessed in vitro and in vivo using confocal laser microscopy and flow cytometry. C. jejuni cells were inactivated with
(w/v) trisodium phosphate (TSP) and the live cells and inactivated cells were distinguished by staining with LIVE/DEAD BacLight Bacteria Viability fluorescent probe. After treatment of TSP for 5 min, most of C. jejuni cells turned to coccoid form from original spiral shape. C. jejuni cells lost total cell viability in the absence of organic nutrients but did not lost total cell viability in the presence of organic nutrients. In vivo test, C. jejuni cells turned to viable but non-culturable (VBNC) form after TSP treatment and remained alive on chicken skin. C. jejuni cells attached on chicken meat would transform to coccoid form by sanitizer treatment, but could possibly be alive by the benefits of organic nutrients present in chicken meat.
Effects of Glucose Degradation Products on Human Peritoneal Mesothelial Cells
Song, Jae-Sook ; Lee, Kyung-Lim ; Ha, Hunjoo ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 308~314
Both high glucose and glucose degradation products (GDP) have been implicated in alterations of peritoneal membrane structure and function during long-term peritoneal dialysis (PD). The present study examined the role of GDP including methylglyoxal (MGO), acetaldehyde, and 3,4-dideoxyglucosone (3,4-DGE) in HPMC activation with respect to membrane hyperpermeability or fibrosis. The role of reactive oxygen species (ROS) and activation of protein kinase C (PKC) in GDP-induced HPMC activation were also examined. Using M199 culture medium as control, growth arrested and synchronized HPMC were continuously stimulated by MGO, acetaldehyde, and 3,4-DGE for 48 hours. Vascular endothelial growth factor (VEGF) was quantified as a marker of peritoneal membrane hyperpermeability and fibronectin and heat shock protein 47 (hsp47) as markers of fibrosis. Involvement of ROS and PKC was examined by the inhibitory effect of N-acetylcystein (NAC) or calphostin C, respectively. MGO significantly increased VEGF (1.9-fold), fibronectin (1.5-fold), and hsp47 (1.3-fold) secretion compared with control M199. NAC and calphostin C effectively inhibited MGO-induced VEGF upregulation. Acetaldehyde stimulated and 3,4-DGE inhibited VEGF secretion. Fibronectin secretion and hsp47 expression in HPMC were not affected by acetaldehyde or 3,4-DGE In conclusion, MGO upregulated VEGF and fibronectin secretion and hsp47 expression in HPMC, and PKC as well as ROS mediate MGO-induced VEGF secretion by HPMC. This implies that PKC activation and ROS generation by GDP may constitute important signals for activation of HPMC leading to progressive membrane hyperpermeability and accumulation of extracellular matrix and eventual peritoneal fibrosis.
Cultural Conditions for Pretense Production by a feather-Degrading Bacterium, Bacillus megaterium F7-1
Son Hong-Joo ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 315~318
The effects of inorganic salts and feather concentrations on pretense production by Bacillus megaterium F7-1 were investigated. Pretense production was dependent on the presence of phosphates in the medium. Supplementation of medium with calcium ion slightly increased protease production. The highest protease production was obtained at
feather. The optimal medium contained
whole feather. By using this optimized medium, increased production of the protease was achieved compared with the cases of using basal medium.
Characterization of Alanine Scanning Mutants of a Peptide Specifically Binding to
Seo, Min-Hee ; Chael, Hee-Kwon ; Myung, Heejoon ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 319~321
We have previously reported the isolation and characterization of peptides binding to
nanoparticles from phage display peptide libraries. One of the peptides (PEP9) was selected and mutant peptide-displaying phages were produced by alanine scanning mutagenesis. The mutant phages were subjected to binding analysis to
nanoparticles. When the proline at residue 4 was substituted by alanine, the binding activity was reduced to
of that of wild type PEP9. Substitution of valine at residue 2, serine at residue 3, and isoleucine at residue 5 also decreased the binding to
. Based on these observations, we concluded that the three dimensional structure generated by residues 2-5 was the critical factor for the binding between PEP9 and the nanoparticle.
Estrogen Effect on the Na,K-ATPase Activity Repressed by IgE-Dependent Histamine-Releasing factor in HeLa Cells
Lee Si-Nae ; Kim Hwa-Jung ; Lee Kyunglim ;
Microbiology and Biotechnology Letters, volume 33, issue 4, 2005, Pages 322~326
IgE-dependent histamine-releasing factor (HRF) is found extracellularly to regulate the degranulation process of histamine in mast cells and basophils and known to play a predominant role in the pathogenesis of chronic allergic disease. HRF has been also identified in the intracellular region of the cell. Previously, we reported that HRF interacts with the 3rd cytoplasmic domain of the alpha subunit of Na,K-ATPase and inhibits Na,K-ATPase activity. Since it is known that estroaen activates the sarcolemmal Na,K-ATPase, we tested whether estrogen recovers the Na,K-ATPase activity repressed by HRF. In this study, we showed that estrogen activates Na,K-ATPase repressed by HRF. RT-PCR and western blot analysis showed that estrogen doesn't reduce the expression level of HRF in HeLa cell, suggesting that this recovery effects of estrogen probably occur via indirect mechanism on HRF and Na,K-ATPase.