• 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.334.1-334.1
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• 2002

Streptomyces scabiei subsp. chosunensis M0137. nonadecanoic acid producer. showed the highest protease activity when grown in OSY medium (oatmeal 1.5%, soybean meal 2%, dried yeast 1 %) supplemented with. glycerol (1 %) and CaCO3 (0.1 %). Two forms of protease(SS-1 and SS-2) were fractionated and purified through Ultrogel AcA 54 gel filtration and DEAE-sepharose CL-6B column chromatography. Both proteases were practically stable in the pH range of 6-10. The optimal pH for the activities of both protease 88-1 and 8S-2 were 7.5 and 8.0. respectively. (omitted)

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2007년도 제48회 학술심포지움 및 추계국제학술대회
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• pp.87.1-87.1
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• 2007

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• 미생물학회지
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• 제46권1호
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• pp.63-67
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• 2010

생리 및 상업적 응용분야에서 중요한 위치를 차지하는 단백질 분해효소는 펩타이드 결합의 가수분해를 촉매한다. 단백질 분해효소를 생산하는 신규 균주를 분리하기 위하여 부패한 나무로부터 균을 분리하여 1% skim milk가 포함된 LB 배지에서 투명환의 생성 능력이 높은 두 균주 WD12와 WD32를 선발하였다. 선발균주의 동정을 위하여 16S rRNA gene의 염기서열을 결정한 후 GenBank 상에 등록된 서열들과 상동성을 조사한 결과, WD12는 Pseudoxanthomonas mexicana와 97.8%, WD32는 99.8%의 상동성을 보여주었다. 계통적 유연관계를 조사한 결과 이들 균들은 P. mexicana와 P. japonensis와 cluster를 형성하였다. WD12와 WD32는 그람 음성 간균으로 catalase와 oxidase 활성을 보였다. WD12의 경우 P. mexicanna와 달리 malate를 동화할 수 있었으며, D-mannose를 동화할 수 없었다. 두 균주의 최적 단백질 분해효소를 생산하는 온도는 $35-37^{\circ}C$로 나타났으며, 최대활성은 WD12가 656 unit/ml, WD32가 267 unit/ml을 나타내었다.

• Journal of Microbiology
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• 제42권2호
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• pp.156-159
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• 2004

In general, the salinity of the ocean is close to 3.5％ and marine vibrios possess the respiratory chain-linked Na$\^$+/ pump. The influence of sodium chloride and the proton conductor carbonylcyanide m-chlo-rophenylhydrazone (CCCP) on the production of extracellular proteases in a marine Vibrio strain was examined. At the concentration of 0.5 M, sodium chloride minimally inhibited the activity of extra-cellular proteases by approximately 16％, whereas at the same concentration, the producton of extra-cellular proteases was severely inhibited. On the other hand, the production of extracellular proteases was completely inhibited by the addition of 2 ${\mu}$M CCCP at pH 8.5, where the respiratory chain-linked Na$\^$+/ pump functions.

• 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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• 제17권5호
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• pp.761-768
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• 2007

Serratia proteamaculans HY-3 isolated from the digestive tract of a spider produces an extracellular protease named arazyme, with an estimated molecular mass of 51.5 kDa. The purified enzyme was characterized as having high activities at wide pH and temperature ranges. We further characterized biochemical features of the enzymatic reactions under various reaction conditions. The protease efficiently hydrolyzed a broad range of protein substrates including albumin, keratin, and collagen. The dependence of enzymatic activities on the presence of metal ions such as calcium and zinc indicated that the enzyme is a metalloprotease, together with the previous observation that the proteolytic activity of the enzyme was not inhibited by aspartate, cysteine, or serine protease inhibitors, but strongly inhibited by 1,10-phenanthroline and EDTA. The araA gene encoding the exoprotease was isolated as a 5.6 kb BamHI fragment after PCR amplification using degenerate primers and subsequent Southern hybridization. The nucleotide sequence revealed that the deduced amino acid sequences shared extensive similarity with those of the serralysin family of metalloproteases from other enteric bacteria. A gene(inh) encoding a putative protease inhibitor was also identified immediately adjacent to the araA structural gene.

• Applied Biological Chemistry
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• 제28권3호
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• pp.209-217
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• 1985

토양으로부터 균체외(菌體外) 단백질 생산균주 17주(株)를 분리하여 이 중 단백질생산이 강한 T219를 선정하여 동정(同定)하였으며 단백질 생산에 영향을 미치는 인자(因子)들을 검토하였다. T219균주(菌株)는 Bacillus속으로 동정(同定)되였으며 균체외(菌體外) 단백질생산 최적온도는 $25^{\circ}C$, pH는 7.5이었다. 단백질생산균주들이 분비하는 단백질에서는 protease와 amylase활성은 보이지 않았으며, 배양시간에 따른 단백질생산은 배양 2일에서 최대로 되었다. yeast extract와 meat extract를 함유한 배지에서 균체증식은 컸지만, 단백질 축적은 거의 없었고, polypeptone은 균체(菌體) 증식과 단백질생산에 큰 효과를 보였다. 또한 배지에 glycine과 L-isoleucine을 혼합하여 첨가하였을때 단백질생산의 효과가 커서 4mg/ml의 단백질축적이 있었다. T219균주(菌株)는 streptemycin의 $30{\mu}g/ml$이상의 농도에서 생육이 저해되었고, EDTA의 5mM농도에서는 생육에 영향을 받지 않았다. 또한, 이들 물질은 단백질생산에 영향을 미치지 않았다.

• Parasites, Hosts and Diseases
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• 제37권1호
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• pp.39-46
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• 1999

As a preliminary study for the explanation of pathobiology of Neodiplostomum seoulense infection. a 54 kDa protease was purified from the crude extract of adult worms by sequential chromatographic methods. The crude extract was subjected to DEAE-Sepharose Fast Flow column, and protein was eluted using 25 mM Tris-HC1 (pH 7.4) containing 0.05. 0.1, 0.2 and 0.4 M NaC1 in stepwise elution. The 0.2 M NaCl fraction was further purified by Q-Sepharose chromatography and protein was eluted using 20 mM sodium acetate (pH 6.4) containing 0.05, 0.1. 0.2 and 0.3 M NaCl, respectively. The 0.1M NaCl fraction showed a single protein band on SDS-PAGE carried out on a 7.5-15% gradient gel. The proteolytic activities of the purified enzyme were specifically inhibited by L-trans-epoxy-succinylleucylamide (4-guanidino) butane (E-64) and iodoacetic acid. The enzyme, cysteine protease. showed the maximum proteolytic activity at pH 6.0 in 0.1 M buffer, and degraded extracellular matrix proteins such as collagen and fibronectin with different activities. It is suggested that the cysteine protease may playa role in the nutrient uptake of N. seoulense from the host intestine.

• 미생물학회지
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• 제39권4호
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• pp.230-234
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• 2003

Bacillus amyloliquefaciens psychrotrophic strain이 분비하는 세포 외 단백질 가수분해효소를 정제하여, endopeptidase 활성에 관한 특성을 분석하였다. Protease SE910로 명명된 효소는 단백질 내부의 leucine에 연결된 peptide 결합만을 가수분해하는 endopeptidase로 작용한다. 효소를 특이한 아미노산 잔기에 작용하는 화학수식제와 반응하였을 때 효소의 활성부위에 관여하는 아미노산 잔기가 수식되었을 때는 효소활성이 저해를 받는다. 본 효소는 serine을 수식하는 PMSF에의해 endopeptidase 활성이 완전히 저해되었으며, 카르복실 기능기를 수식하는 화학수식제에 의해 저해되었고, lysine을 화학수식하는 PLP에 의해서는 큰 영향을 받지 않았다. 이것은 본 효소의 endopeptidase 활성에 serine과 aspartic acid 잔기가 관여하는 것을 의미한다. 구조적으로 leucine을 포함하는 유도체인 bestatin은 효소의 endopeptidase 활성을 경쟁적으로 저해하였다.

• 한국발생생물학회지:발생과생식
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• 제14권3호
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• pp.207-214
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• 2010

The tremendous changes of uterine endometrium are observed during early pregnancy and protease and their inhibitors are involved in regulation of cell proliferation and remodeling of the tissues through remodeling the extracellular matrix (ECM). Some of the proteases and protease inhibitors have been suspected to a factor in endometrial changes but many parts of their expression profiles and the physiological roles are not uncovered. To evaluate the functional roles of them, in this study the expression profiles of proteases and protease inhibitors were analyzed using real-time quantitative PCR analysis. Mmp9 (matrix metalloproteinase 9) mRNA levels peaked on day 4 at the time of implantation. On the other hand, Ela2 (neutrophil elastase, NE) mRNA levels were peaked on day 2 of pregnancy. Its expression were decreased until day 4 of pregnancy but increased rapidly until day 7 of pregnancy and decreased again. NE inhibitor Slpi (secretory leukocyte protease inhibitor, SLPI) mRNA levels were related with the implantation stage and with the levels of Ela2. At the time of implantation the expression levels of Slpi mRNA were about 5 times higher than the Ela2 mRNA in the uterus. In the implantation stage embryos, Mmp9 specific mRNA was only detected at the blastocyst. On the other hand, the expression level of SLPI was higher than that of the Ela2 mRNA at blastocyst and 4.5 day p.c. embryos. Based on these results it is suggested that MMP9, SLPI, and NE have important physiological role in embryo implantation both in uterus and embryos.