• Journal of Soil and Groundwater Environment
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• v.14 no.6
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• pp.45-52
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• 2009

Naphthalene-degrading bacteria Pseudomonas aeruginosa CZ6 isolated from contaminated soil can adhere to crystal naphthalene and produce extracellular polymeric substance. LB, YM and MSM medium were used as culture mediums to investigate the formation of biofilm. Biofilm was developed the most in LB medium by Pseudomonas aeruginosa CZ6. In the culture, strain CZ6 growth was rarely affected by naphthalene concentration. Optimal culture condition was $30^{\circ}C$ and pH 7 at 0.10% substrate and 150 rpm shaking. The effect of culture medium on naphthalene degradation in the two soil slurry system was evaluated. The initial degradation rate of naphthalene was highest in the MSM medium of soil slurry. However, the sorbed naphthalene was rapidly degraded at the LB medium when naphthalene availability in liquid was limited. The results of this study suggest that biofilm formation and extracellular polymeric substance production increased bioavailability of soil sorbed naphthalene.

• 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.

• Journal of Microbiology and Biotechnology
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• v.14 no.2
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• pp.430-434
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• 2004

Three extracellular protease-deficient Bacillus subtilis strains were transformed with the plasmid pCK98 containing the endo-$\beta$-1,4-glucanase (Eng) gene of B. subtilis BSE616. The three transformants, B. subtilis DB104 (pCK98), WB600 (pCK98) and WB700 (pCK98), produced the same high level of enzyme activity and showed similar patterns of cell growth and enzyme production. When B. subtilis DB 104 (pCK98), a two-extracellular protease deficient strain, was cultured for 22 h, almost all the secreted enzyme was found to be in the completely cleaved form by both activity staining and Western blotting studies. B. subtilis WB600 (pCK98), a six-extracellular protease-deficient strain, produced a partially cleaved form in addition to the intact form of the enzyme, although the degree of internal cleavage of the enzyme was greatly reduced. With B. subtilis WB700 (pCK98), a seven-extracellular protease-deficient strain, almost all the enzyme was produced as the intact uncleaved form. This study illustrates that a role of the V pr protease is to degrade foreign proteins produced in B. subtilis and WB700 is a suitable expression system for producing the intact form of the Eng and other foreign proteins that may lose at least part of their efficacy due to internal proteolytic cleavage.

• Current Research on Agriculture and Life Sciences
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• v.31 no.1
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• pp.30-37
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• 2013

Bacillus subtilis and Bacillus amyloliquefaciens are closely related species that share a similar genomic background, and are both known to secrete large amounts of proteins directly into a medium. The extracellular proteomes of two strains of Bacillus subtilis and two strains of Bacillus amyloliquefaciens were compared by 2-D gel electrophoresis during the late exponential growth phase. The relative abundance of some minor protein spots varied among the four strains of Bacillus. Over 123 spots of extracellular proteins were visualized on the gel for B. subtilis CH 97, 68 spots for B. subtilis 3-5, 230 spots for B. amyloliquefaciens CH 51, and 60 spotsfor B. amyloliquefaciens 86-1. 2D gel electrophoresis images of the four Bacillus strains showed significantly different protein profiles. Consistent with the 2D gel electrophoretic analysis, most of the B. subtilis proteins differed from the proteases secreted by the B. amyloliquefaciensstrains. Among the proteins identified from B. subtilis, approximately 50% were cytoplasmic and 30% were canonically extracellular proteins. The secreted protein profiles for B. subtilis CH 97 and B. subtilis 3-5 were quite different, as were the profiles for B. amyloliquefaciens CH 51 and 86-1. The four proteomes also differed in the major protein composition. The B. subtilis CH 97 and B. amyloliquefaciens CH 51 proteomes both contained large amounts of secreted hydrolytic enzymes. Among the four strains, B. subtilis 3-5 secreted the least number of proteins. Therefore, even closely related bacteria in terms of genomic sequences can still have significant differences in their physiology and proteome layout.

• Korean Journal of Microbiology
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• v.36 no.1
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• pp.64-68
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• 2000

The immunopotentiating effects of the polysaccharides, both intracellular and extracellular, was examined by an animal feeding test. The results are summarized as follows. The oral administration of intracellular and extracellular polysaccharides of Agrocybe cylindracea for 10 days resulted in the enhanced phagocytic activity of peritoneal exudate cells(PEC), spleen cells(SC), and monolymphocytes(ML). In the experiment of PFC(plaque forming cell) and RFC(rosette forming cell), the results showed that all the polysaccharide fractions enhanced the immune related cells. The EAC II group(the extracellular polysaccharide of Agrocybe cylindracea 10 mg/0.2 ml distilled water/day/ mouse) increased the PFC and RFC by 46~50% and 43%, respectively, compared to the control group. On the other hand, the IAC I group(the intracellular polysaccharide of Agrocybe cylindracea 1 mg/0.2 ml distilled water/day/mouse) increased the PFC and RFC by 49~70% and 91%, respectively. In terms of the mitogenic activity, the extracellular polysaccharides of A. cylindracea showed a higher activity than the intracellular polysaccharides.

• The Journal of the Korean dental association
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• v.55 no.1
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• pp.30-41
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• 2017

Inadequate keratinized mucosa around dental implants can lead to more plaque accumulation, tissue inflammation, marginal recession and attachment loss. We evaluated the effects of free gingival and extracellular matrix membrane grafts performed to increase the insufficient width of keratinized tissue around dental implants in the posterior mandible. A 47-year-old female patient presented with discomfort due to swelling of the lower right second premolar area. Due to severe destruction of alveolar bone, the tooth was extracted. After 3 months, a guided bone regeneration (GBR) procedure was performed and then a dental implant was placed 6 months later. During the second-stage implant surgery, free gingival grafting was performed to increase the width of the keratinized tissue. After 12 months, a clinical evaluation was performed. A 64-year-old female patient had a missing tooth area of bilateral lower molar region with narrow zone of keratinized gingiva and horizontal alveolar bone loss. Simultaneous implant placement and GBR were performed. Five months after the first-stage implant surgery, a gingival augmentation procedure was performed with an extracellular matrix membrane graft to improve the width of the keratinized tissue in the second-stage implant surgery. After 12 months, a clinical evaluation was performed. In these two clinical cases, 12 months of follow-up, revealed that the increased width of the keratinized tissue and the deepened oral vestibule was well maintained. A patient showed a good oral hygiene status. In conclusion, increased width of keratinized tissue around dental implants could improve oral hygiene and could have positive effects on the long-term stability and survival rate of dental implants. When planning a keratinized tissue augmentation procedure, clinicians should consider patient-reported outcomes.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.93-98
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• 1993

To investigate the organic matter transformation in aquatic environment, seasonal fluctuations of heterotrophic activity and microbia] extracellular enzyme activity were studied in Paldang Lake, Korea. The turnover time in the water column and the sediment at the station I fluctuated between 3 -I ,300 hrs and 17-170 hrs for glucose, 5 -1.900 hrs and 15-240 hrs for protein hydrolysate and 4-350 hrs and 15-230 hrs for acetic acid, respectively, indicating that the seasonal turnover time of organic substrates fluctuated drastically. The respiration ratios of glucose. protein hydrolysate and acetate were 23-32%, 38-41% and 22-28% in the water column and 34%, 61% and 41% in the sediment. respectively. These results showed that the respiration ratios in the sediment were higher than those in the water column regardless of kinds of organic substrates. The bacterial extracellular enzyme activities of $\alpha$-glucosidase. $\beta$-glucosidase, N-acetyl-$\beta$-D-glucosaminidase and aminopeptidase were 32-44%. 31-32%, 18-34% and 61-67% in the water column, and 34%. 40%, 23% and 65% in the sediment. respectively.

• Journal of Microbiology and Biotechnology
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• v.20 no.10
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• pp.1378-1385
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• 2010

A novel agarolytic bacterium, KY-YJ-3, producing extracellular agarase, was isolated from the freshwater sediment of the Sincheon River in Daegu, Korea. On the basis of Gram-staining data, morphology, and phylogenetic analysis of the 16S rDNA sequence, the isolate was identified as Cellvibrio sp. By ammonium sulfate precipitation followed by Toyopearl QAE-550C, Toyopearl HW-55F, and MonoQ column chromatographies, the extracellular agarase in the culture fluid could be purified 120.2-fold with a yield of 8.1%. The specific activity of the purified agarase was 84.2 U/mg. The molecular mass of the purified agarase was 70 kDa as determined by dodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal temperature and pH of the purified agarase were $35^{\circ}C$ and pH 7.0, respectively. The purified agarase failed to hydrolyze the other polysaccharide substrates, including carboxymethyl-cellulose, dextran, soluble starch, pectin, and polygalacturonic acid. Kinetic analysis of the agarose hydrolysis catalyzed by the purified agarase using thin-layer chromatography showed that the main products were neoagarobiose, neoagarotetraose, and neoagarohexaose. These results demonstrated that the newly isolated freshwater agarolytic bacterium KY-YJ-3 was a Cellvibrio sp., and could produce an extracellular ${\beta}$-agarase, which hydrolyzed agarose to yield neoagarobiose, neoagarotetraose, and neoagarohexaose as the main products.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1993-2005
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• 2016

Bacillus pumilus is one of the most characterized microorganisms that are used for high-level production of select industrial enzymes. A novel B. pumilus SCU11 strain possessing high alkaline protease activity was obtained in our previous work. The culture supernatant of this strain showed efficient dehairing capability with minimal collagen damage, indicating promising potential applications in the leather industry. In this study, the strain's extracellular proteome was identified by LC-MS/MS-based shotgun proteomic analysis, and their related secretory pathways were characterized by BLAST searches. A total of 513 proteins, including 100 actual secreted and 413 intracellular proteins, were detected in the extracellular proteome. The functions of these secreted proteins were elucidated and four complete secretory systems (Sec, Tat, Com, and ABC transporter) were proposed for B. pumilus. These data provide B. pumilus a comprehensive extracellular proteome profile, which is a valuable theoretical and applicative basis for future genetic modifications and development of industrial enzymes.

• The Korean Journal of Physiology
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• v.24 no.2
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• pp.281-291
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• 1990

The $Ca^{2+}-substitutional$ roles of strontium for the contractile processes were investigated in the rabbit renal artery. The contractions induced by either norepinephrine or high $K^+$ in the condition which intra- and extracellular $Ca^{2+}$ were replaced by $Sr^{2+}$, i.e. $Sr^{2+}-mediated$ contractions, were dose-dependent. And then the maximal amplitude of contraction, as compared with $Ca^{2+}-mediated$ contraction, was about 50% in norepinephrine and about 70% in high $K^+$. The $Sr^{2+}-mediated$ contractions were independent in the contraction by norepinephrine $(10^{-5}M)$ but dependent in those by high $K^+(100\;mM)$ on the extracellular $Sr^{2+}$ concentration. Also $Sr^{2+}-mediated$ contractions induced by norepinephrine were observed in the $Sr^{2+}-free$ Tyrode's solution. The $Sr^{2+}-mediated$ contractions induced by either norepinephrine or high $K^+$ were suppressed by verapamil, a $Ca^{2+}-channel$ blocker. By extracellular addition of $Sr^{2+}$, the $Ca^{2+}-mediated$ contractions induced by norepinephrine $(10^{-5}M)$ or 40 mM $K^+$ were inhibited but those by high $K^+(100\;mM)$ were increased. And the $Sr^{2+}-mediated$ contractions were increased by extracellular addition of $Ca^{2+}$ but did not reach the level of $Ca^{2+}-mediated$ contraction. Therfore it is suggested that in the vascular smooth muscle of rabbit renal artery $Sr^{2+}$ could enter the smooth muscle cells easily through the potential-operated calcium channel (POC) but not easily through the receptor-operated calcium channel (ROG), and $Sr^{2+}$ might be stored in the intracellular $Ca^{2+}-binding$ site and released by NE and induced the contraction by a way of activating directly the contractile apparatus.