• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.332.2-333
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• 2002

Three alkaline proteases. designated JB-1. JB-2, and JB-3, are extracellular enzymes produced by Bacillus spp. JB411 which was isolated korean soil. They were separated by DEAE-sepharose CL-6B gel. and further purified using ammonium sulfate precipitation. ultra membrane filtration. and Ultrogel AcA gel filtration. The optimun pH values of proteases IB-1. JB-2. and JB-3. were shown to be 9.5. 9.5 and 7.5. respectively. All three proteases were stable in the pH range of 5-11. (omitted)

• 미생물학회지
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• 제27권3호
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• pp.245-249
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• 1989

Bacillus licheniforntis에서 분비된 단백질 추출물에서 ammonium sulfate fractionation과 Sephadex G-75젤 여과 크로마토그라피를 사용하여 부분 정제된 2개의 서로 다른 활성을 갖는 단백질 분해 효소를 얻었다. 단백질 분해 효소 type l은 카제인에 대해 비교적 높은 활성을 보이며 organotluoride와 EOTA에 의한 억제에 매우 민감했다 이 효소의 최적 pH는 7.5였으며 $75^{\circ}C$에서 빠르게 변성되었다. 단백질 분해효소 type II는 type I에 비해 낮은 카제인 분해 활성을 보이며 ImM PMSF와 10mM EOTA용액과 함께 $37^{\circ}C$에서 30분 간 반응시키면 역사 활성이 억제되었다 한편 이 효소 는 pH 8.0에서 최대 활성을 나타내고 $71^{\circ}C$ 에서 상대적으로 늦게 불환성화 되었다.

• Journal of Microbiology and Biotechnology
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• 제16권1호
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• pp.5-14
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• 2006

Streptomycetes are Gram-positive microorganisms producing secondary metabolites through unique physiological differentiation [4]. The microbes show unusual morphological differentiation to form substrate mycelia, aerial mycelia, and arthrospores on solid medium [19]. Substrate mycelium growth is sustaining with sufficient nutrients in the culture medium. The concentration of a specific individual substrate in the culture environment is the most important extracellular factor allowing vegetative mycelia growth, where extracellular hydrolytic enzymes participate in the utilization of waterinsoluble substrates. However, with starvation of nutrients in the culture medium, the vegetative mycelia differentiate to aerial mycelia and spores. It has been considered that shiftdown of essential nutrients for mycelia growth is the most important factor triggering morphological and physiological differentiation in Streptomyces spp. Since proteineous macromolecule compounds are the major cellular components, these are faced to endogenously metabolize following a severe depletion of nitrogen source in culture nutrients (Fig. 1). Various proteases were identified of which production was specifically related with the phase of mycelium growth and also morphological differentiation. The involvement of proteases and protease inhibitor is reviewed as a factor explaining the mycelium differentiation in Streptomyces spp.

• 미생물학회지
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• 제33권2호
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• pp.136-141
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• 1997

한국의 전통 발효식품인 된장에서 단백질 분해효소를 분비하는 미생물의 존재를 규명하기 위해, skim milk 한천배지를 이용하여 투명환을 보이는 균주들을 얻었고, 각종 생리 및 생화학적 검사, VITEK system, MIDI system을 통해 이들 중, JH-1, SH-5, SH-7의 3 strain이 Bacillus cereus임을 동정하였다. JH-1과 SH-5 균주가 분비하는 단백질 분해효소는 pH 9와 40^{\circ}C.$에서 최적활성을 보였으며, SH-7 균주는 pH 8과 50^{\circ}C.$에서 최적활성을 보였다. 또한 이들 3균주는 blood agar plate에서 용혈능력을 보였다.

• 한국균학회지
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• 제36권2호
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• pp.178-182
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• 2008

콘텍트렌즈의 착용이 증가하면서 알칸사아메바에 의한 각막염이 급증하지만 아직도 이병을 치료하기가 매우 어려운 실정이다. 이 아메바에서 나오는 serine 단백질분해효소는 각막염의 유발에 중요한 역할을 하고 있다고 알려졌다. 본 연구에서는 A. castellanii의 세포외 serine 단백질분해효소의 활성을 저해하는 능력이 우수한 추출물을 230종 버섯의 메탄올 추출물로부터 선별하였다. 버섯을 메탄올과 물($65^{\circ}C$)로 추출한 후 여과.농축하여 이 추출물을 0.3 mg/ml의 농도로 효소와 반응시킨 후 기질 N-succinylala-ala-pro-phe p-anilide을 넣고 효소의 활성을 측정하여 우수한 저해능을 가진 추출물을 선별하였다. 이렇게 선별된 추출물을 가지고 성질을 연구하였는데, 한 추출물로 활성이 완전하게 억제되지 않았으나 두 추출물을 조합하여 사용하면 활성을 완전하게 억제할 수 있었다. 이 추출물은 다른 serine 단백질분해효소인 인간과 소의 트롬빈과 일반적인 단백질분해효소인 Protease K에는 영향을 미치지 않았다.

• 대한약침학회지
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• 제18권2호
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• pp.19-25
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• 2015

Objectives: Capsaicin (CAP) is the chief pungent principle found in the hot red peppers and the chili peppers that have long been used as spices, food additives and drugs. This study investigated the anticancer potential of CAP through its ability to modify extracellular matrix components and proteases during mice lung carcinogenesis. Methods: Swiss albino mice were treated with benzo(a) pyrene (50 mg/kg body weight dissolved in olive oil) orally twice a week for four successive weeks to induce lung cancer at the end of $14^{th}$ week. CAP was administrated (10 mg/kg body weight dissolved in olive oil) intraperitoneally. Extracellular matrix components were assayed; Masson's trichome staining of lung tissues was performed. Western blot analyses of matrix metalloproteases 2 and 9 were also carried out. Results: In comparison with the control animals, animals in which benzo(a)pyrene had induced lung cancer showed significant increases in extracellular matrix components such as collagen (hydroxy proline), elastin, uronic acid and hexosamine and in glycosaminoglycans such as hyaluronate, chondroitin sulfate, keratan sulfate and dermatan sulfate. The above alterations in extracellular matrix components were effectively counteracted in benzo(a)pyrene along with CAP supplemented animals when compared to benzo(a) pyrene alone supplemented animals. The results of Masson's trichome staining for collagen and of, immunoblotting analyses of matrix metalloproteases 2 and 9 further supported the biochemical findings. Conclusion: The apparent potential of CAP in modulating extracellular matrix components and proteases suggests that CAP plays a chemomodulatory and anti-cancer role working against experimentally induced lung carcinogenesis.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2005년도 International Meeting of the Microbiological Society of Korea
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• pp.226.3-226
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• 2005

• 셀메드
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• 제4권1호
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• pp.6.1-6.12
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• 2014

Anthrax is the deadly disease for human being caused by Bacillus anthracis. Instantaneous research work on the mode of infection of the organism revealed that different proteases are involved in different steps of pathogenesis. Present study reports the in silico characterization and the detection of pathogenic proteases involved in anthrax infection through protein-protein interaction. A total of 13 acid, 9 neutral, and 1 alkaline protease of Bacillus anthracis were selected for analysing the physicochemical parameter, the protein superfamily and family search, multiple sequence alignment, phylogenetic tree construction, protein-protein interactions and motif finding. Among the 13 acid proteases, 10 were found as extracellular enzymes that interact with immune inhibitor A (InhA) and help the organism to cross the blood brain barrier during the process of infection. Multiple sequence alignment of above acid proteases revealed the position 368, 489, and 498-contained 100% conserved amino acids which could be used to deactivate the protease. Among the groups analyzed, only acid protease were found to interact with InhA, which indicated that metalloproteases of acid protease group have the capability to develop pathogenesis during B. anthracis infection. Deactivation of conserved amino acid position of germination protease can stop the sporulation and germination of B anthracis cell. The detailed interaction study of neutral and alkaline proteases could also be helpful to design the interaction network for the better understanding of anthrax disease.

• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.639-644
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• 1998

Extracellular serine proteases were isolated from a soil bacterium, alkalophilic coryneform bacterium TU-19, which have been grown in a liquid medium optimized at 3$0^{\circ}C$ and pH 10.0. Three different sizes, 120 kDa (protease I), 80 kDa (protease II), and 45 kDa (protease III), of serine pro teases were purified using Sephadex G-150 and QAE-Sephadex chromatography (Kang et al. 1995. Agric. Chem Biotech. 38: 534-540). SDS-PAGE showed that the 120 kDa protease was degraded into the 80 kDa protease in 20 mM Tris-HCI (pH 8.0) buffer solution. This degradation was enhanced in the presence of 0.5 M NaCl and 5 mM EDTA, but was inhibited in the presence of 5 mM $CaCl_2$. These results indicated that the $Ca^{2＋}$ ion seems to stabilize the 120 kDa protease like other proteases derived from Bacillus species. The $NH_2$-terminal amino acid sequences of the 10 residues of both proteases were completely identical: Met-Asn-Thr-Gln-Asn-Ser-Phe-Leu-Ile-Lys. In contrast to this, the 80 kDa protease has 1.5 times higher specific activity than the 120 kDa protease does (Kang et al. 1995. Agric. Chern. Biotech. 38: 534-540). Therefore the C-terminal of the 120 kDa protease seems to be autolyzed to the 80 kDa protease but this autolysis did not decrease the protease activity. Optimum pH and temperature of both 80 kDa and 120 kDa proteases were pH 10.5 and $45^{\circ}C$, respectively, and pH and thermal stability were almost identical. Several divalent ions except the $Fe^{2＋}$ ion showed similar effects on activities of both proteases, which are similarly resistant to three different detergents.

• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.457-464
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• 2006

Three extracellular proteases (Vpr, peptidase T, and subtilisin) were identified from the culture supernatant of Bacillus subtilis KCTC 3014. All the proteins were partially purified as a mature form by using a DEAE-cellulose ion-exchange column chromatography. Their activities were determined by using zymography and densitometry. The relative molecular masses of Vpr and peptidase T (PepT) were determined to be 68 and 48 kDa by SDS-PAGE and zymography, respectively. However, subtilisin formed a 'binding mode' at the top of the separating gel. After denaturation by boiling at $100^{\circ}C$ for 5 min, its molecular mass was determined to be 29 kDa, whereas its activity was lost. The optimal pH of Vpr, PepT, and subtilisin were 9.0, 6.0-7.0, and 7.0-8.0, respectively. The optimal temperature of Vpr, PepT, and subtilisin was 40, 50, and $40^{\circ}C$, respectively. Inhibitor test revealed that Vpr and subtilisin were serine proteases and that PepT was a metalloprotease. Interestingly, we found that Vpr showed no enzyme activity on a 2DE zymogram gel. Three genes, vpr, pepT, and apr (encoding subtilisin protein), were cloned and their nucleotide and deduced amino acid sequences were determined.