• 한국지하수토양환경학회지:지하수토양환경
• /
• 제14권6호
• /
• pp.45-52
• /
• 2009

나프탈렌 분해균주인 Pseudomonas aeruginosa CZ6을 오염된 토양에서 분리하였으며 분리된 균주는 결정상태의 나프탈렌에 부착하고 그 주변에 extracellular polymeric substance를 분비하는 특성을 가졌다. LB, YM과 MSM 배지를 사용하여 배지의 종류에 따른 biofilm 생성량을 측정한 결과, LB 배지에서 biofilm이 가장 많이 생성되는 것으로 나타났다. 나프탈렌을 기질로 한 배양조건에서 균주는 기질의 농도 영향을 크게 받지 않고 0.10% 기질, 150 rpm 조건에서 최적 배양조건은 $30^{\circ}C$, pH 7로 나타났다. 두 가지 토양에서 배지의 종류에 따른 나프탈렌의 분해특성을 관찰한 결과 초기에는 MSM 배지에서 나프탈렌이 가장 많이 분해가 되었다. 그러나 생물이용성이 제한을 받는 조건에서 LB 배지의 나프탈렌이 가장 빨리 제거가 되었다. 이런 결과는 biofilm의 형성과 extracellular polymeric substance 생성이 토양에 흡착된 잔류 나프탈렌의 생물학적 이용성을 향상시키기 때문인 것으로 사료된다.

• 미생물학회지
• /
• 제44권1호
• /
• pp.1-9
• /
• 2008

본 논문은 벼 흰 잎마름병균인 Xanthomonas oryzae pv. oryzae(Xoo)의 병원성 유전자의 분자유전학적 연구현황을 기술하고자 한다. 또한 국내 고유 벼 흰 잎마름병균 KACC10331의 유전체해독 정보를 기반으로 다른 Xanthomonas의 유전체와 비교 분석함으로써, Xoo의 주요 병원성 유전자의 분자구조를 구명하고자 한다. 이를 통해 Xoo 고유 병원성 유전자 탐색 및 기능 해석을 위한 기초자료를 제공하는데 목적이 있다. Xoo 유전체에는 5개 과(family)에 속하는 9 type의 Insertion sequence(IS)가 611 copy로 존재하고 있으며, 주로 병원성 관련 유전자군 주위에 많이 분포하고 있는 것으로 나타났다. 현재까지 연구가 수행된 주요 병원성 관련 유전자인 hypersensitive response and pathogenicity (hrp) 유전자, extracellular polysaccharide (EPS) 유전자, extracellular enzyme 유전자, lipopolysaccharides (LPS) 유전자, 그리고 avilulence 유전자의 분자유전학적 연구현황을 기술하였다.

• Journal of Microbiology and Biotechnology
• /
• 제14권2호
• /
• pp.430-434
• /
• 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
• /
• 제31권1호
• /
• pp.30-37
• /
• 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.

• 미생물학회지
• /
• 제36권1호
• /
• pp.64-68
• /
• 2000

Agrocybe cylindracea으로부터 분리 배양한 균사체 배양액을 이용하여 균사체내 다당류(IP)와 균사체외 다당류(EP)를 분리하고, 동물실험을 통한 각 다당류의 면역증강활성에 관해 실험을 실시하였다. 탐식능에 대한 실험결과, IP와 EP 모두는 복강 침출 세포(PEC)에서 대조군에 비해 6.6-50%, 말초 임파구(ML)에서 67-167%, 비장(SP)에서 16-90%의 전반적으로 높은 탐식능을 나타냈다. 또한 항체 생성능 측정을 위한 plaque forming cell(PFC)와 rosette forming cell(RFC)의 실험결과에서 볼 때, PFC형성능은 대조군에 비해 EP투여 II군에서 46-50%, IP투여 I군에서 49-70%의 활성증가를 나타내었다. 또한 RFC형성능은 대조군에 비해 IP투여 I군에서 91%의 증가를, EP투여 I군에서 33%, EP투여 II군에서 43%의 활성증가를 나타내어 높은 항체생성이 기대되었다. Mitogen 활성은 농도 의존적으로 세포 증식능을 보였으며, IP와 EP를 비교할 때, EP는 IP에 비해 약2.5배의 세포증식 활성을 보였다.

• 대한치과의사협회지
• /
• 제55권1호
• /
• pp.30-41
• /
• 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.

• 미생물학회지
• /
• 제31권1호
• /
• pp.93-98
• /
• 1993

수계생태계에서 유기물질의 순환을 이해하기 위하여 팔당호에서 종속영양 활성도와 세균세포의 효소활성의 계절절 변화를 연구하였다. 팔당호 I 의 glucose 전환시간은 수층, 퇴적토에서 2-1,300 시간, 17-170 시간, protein hydrolysate 는 5-900 시간, 15-240 시간, acetic acid 는 4-350 시간, 15-230 시간으로 계절적인 변화를 나타냈다. Glucose, protein hydrolysate, acetate 각각의 호흡율은 수층에서 23-32%, 38-41%, 22-28%로 나타났고 퇴적토에서는 34%, 61% and 41% 로 나타났다. 이 결과로 3가지 유기물질 종류 모두가 수층보다 퇴적토에서는 높은 율로 호흡됨을 알 수 있었다. 한편 세균의 $\alpha$-glucosidase, $\beta$-glucosidase, N-acetyl-$\beta$-D-glucosaminidase, aminopeptidase 활성력을 살펴본 결과 수층에서는 효소 각각에 대해 32-44%, 31-32%, 18-34% 61-67% 의 범위를 나타내었고 퇴적토에서는 34%, 40%, 23% 65%로 나타났다.

• Journal of Microbiology and Biotechnology
• /
• 제20권10호
• /
• pp.1378-1385
• /
• 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
• /
• 제26권11호
• /
• pp.1993-2005
• /
• 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
• /
• 제24권2호
• /
• pp.281-291
• /
• 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.