• 생명과학회지
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• 제6권4호
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• pp.241-249
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• 1996

Vivrio mimicus (ATCC 33568)의 최적 배양조건하에서 배양액으로부터 collagenase를 황산암모늄 염석과 DEAE-Sephadex A-50 이온코환크로마토그래피에 의해 분리. 정제하였다. SDS-PAGE 전기영동분석법 및 겔여과법으로 정제된 collagenase의 분자량은 42 kDa 이였다. 기질인 불용성 콜라겐(Type I)에 대한 collagenase의 최적 pH 및 온도는 각각 7.75 및 28$\circ$였다. 금속착물제와 serine protease 저해제는 collagenase의 활성을 저해하였지만 L-cysteine과 histidine은 효소의 활성을 저해하지 않았다. collagenase의 아미노산 조성은 glycine 및 alanine의 아미노산 잔기가 많이 함유되어 있었다. 불용성 (Type I) 콜라겐에 대한 collagenase의 속도상수인 K$_{m}$ 값 및 R$_{cat}$/K$_{m}$값은 각각 2.86 mg/ml 및 972.28 U/mg-protein 이었다.

• Journal of Microbiology and Biotechnology
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• 제8권3호
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• pp.245-250
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• 1998

A gene encoding an extracellular collagenase from the marine bacterium Vibrio vulnificus CYK279H was cloned into E. coli JM83 using the multicopy plasmid vector pUC19. The cloned strain of recombinant E. coli showing collagenase activity had an insert fragment of 3.5 kb and was named E. coli JM83/pKCL 279H. The cloned strain produced two different collagenase during cultivation. These enzymes, named collagenase-I and -II, were purified from the culture supernatant. SDS-PAGE indicated that collagenase-I had a molecular weight of 41 kDa and collagenase-II had a weight of 37 kDa. The N-terminal amino acid sequence of collagenase-I from the cloned strain, E. coli JM83/pKCL279H was determined and was not found to be similar to any other known collagenases. The optimum pH and temperature of the purified collagenase-I were 7.8 and $37^{\circ}C$, respectively.

• 생명과학회지
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• 제15권4호
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• pp.657-663
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• 2005

Collagnase는 천연 collagen의 triple-stranded helix를 분해할 수 있는 protease로서 조직의 수복과 재생 과정에서 collagen의 재형성과 세포의 이동에 아주 중요한 역할을 하고, 숙주 감염시에는 collagen 기질을 빠르게 분해함으로써 감염을 돕는다. 본 연구에서는 일반가정에서 식용하는 김치로부터 collagenase를 생산하는 균주를 분리하여 Bacillus subtilis로 동정하였으며 이를 Bacillus subtilis JS-17이라 명명하였다. Bacillus subtilis JS-17이 생산하는 collagenase의 최적 생산 조건은 $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4{\cdot}4H_2O,\;0.1\%\;citrate,\;0.1\%\;CaCl_2(pH\;7.0)$의 배지에서 $30^{\circ}C$, 200 rpm으로 72시간 동안 배양하는 것이다. 최적 조건에서 Bacillus subtilis JS-17이 생산하는 collagenase를 Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography, DEAE-Sephadex A-30 column chromatography를 거쳐 분리 정제하고, 얻어진 정제 효소의 특성에 대하여 검토하였다. 정제된 collagenase의 비활성은 growth medium에서 192.1 units/mg였고, $1.1\%$의 수율로 얻어졌으며 분자량은 28 kDa이었다. 정제된 collagenase는 $55^{\circ}C$까지는 $100\%$의 활성을 유지하였고 $65^{\circ}C$에서도 $60\%$ 정도의 활성을 유지하였다. 또한 pH $6.0\~9.8$에서 $60\%$ 이상의 활성을 유지하였다. 정제된 collagenase는 metalloprotease inhibitor인 EDTA와 O-phenanthroline에 의해 효소 활성이 감소하였을 뿐만 아니라 Ammoninum persulfate, L-cysteine, N-ethylmaleimide, SDS, $NaN_3$, NaF, $KMnO_4$, PMSF에 대해서도 활성이 감소하였다. 정제된 collagenase를 여러 가지 기질에 대해 효소 활성을 비교한 결과 collagen (type I)에 대해 기질 특이성을 가지고 있었다.

• 한국수산과학회지
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• 제44권5호
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• pp.456-463
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• 2011

We attempted to isolate a collagenase-1 inhibitory peptide from skate Dipturus chilensis skin protein. The protein from skate skin was digested by various enzymes (alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin) to produce a collagenase-1 inhibitory peptide. The collagenase-1 inhibitory activity of the peptides obtained was measured by gelatin digestion assay. Among the six hydrolysates, pepsin hydrolysate exhibited the highest collagenase-1 inhibitory activity. The peptide showing strong collagenase-1 inhibitory activity was purified by Sephadex G-25 gel chromatography and HPLC using an octadecylsilyls (ODS) column. The amino acid sequence of purified collagenase-1 inhibitory peptide was identified to be Asn-Leu-Asp-Val -Leu-Glu-Val-Phe (961 Da) by quadrupole time of flight (Q-TOF) and electrospray ionization mass spectrometry (ESI-MS) mass spectroscopy. The $IC_{50}$ value of purified peptide was 87.0 ${\mu}M$. Moreover, the peptide did not exhibit cytotoxic effects on human dermal fibroblast cell lines.

• 대한치과교정학회지
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• 제20권2호
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• pp.227-245
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• 1990

The purpose of this study was to analyze the reorganization of periodontal ligament after collagenase treatment with autoradiography. The author compared the collagenase-treated experimental group and no-treated experimental group with control group. Fourty eight Sprague-Dawley rats were divided into nine groups, including normal control and immediate group. Closed coil springs were used between the upper incisors and the first molars with 100 grams. Collagenase and $^3H-proline$ were adminstered and the samples were sacrificed and sectioned. After being dipped into the NTB-3 emulsion the samples were analyzed with light microscope under H/E stain. Data were analyzed by t-test and ANOVA. The results were as follows: 1) Generally collagenase-treated groups got more $^3H-proline$ uptake than no-treated groups. 2) Compared with normal control group, collagenase-treated group had the same $^3H-proline$ uptake in amount at 21th day. 3) Among cemento-enamel junction, middle, apex areas, cementa-enamel junction area of collagenase-treated group arrived at normal control level earlier than no-treated group. 4) Cemento-enamel junction area had the most $^3H-proline$ incorporation amount in no-treated group, but apex area had the most in collagenase group.

• 한국어병학회지
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• 제12권2호
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• pp.83-88
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• 1999

녹차 추출물을 이용하여 각종 어병세균의 collagenase 활성과 생성 억제 및 생육저해 효과를 조사하였다. 어병세균의 감염시에 필요한 효소인 collagenase활성을 8종의 어병세균에 대하여 측정한 결과 0.08-0.7unit/ml 범위로 나타났다. 그중 collagenase활성이 높은 Edwardsiella tarda의 collagenase 활성은 0.2 mg/ml의 녹차 추출물이 첨가된 반응액에서 거의 100% 저해되었다. E. tarda의 collagenase 생합성은 ST배지에서 $25^{\circ}C$, 16시간 배양하였을 때 가장 높게 나타났다. 이 효소는 0.08 mg/ml의 녹차 추출물이 첨가된 배양액에서 1/3이상 생합성이 감소하였다. 또한 균의 생육에 미치는 녹차 추출물의 영향을 조사한 결과 8 mg/ml이 첨가된 배양액 내에서 균의 생육이 완전히 억제되었다.

• Applied Biological Chemistry
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• 제51권3호
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• pp.164-170
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• 2008

Coenzyme $Q_{10}$과 6가지 coenzyme Qn 유토체들을 합성하였다. Coenzyme Qn유도체들을 murine melanoma(B16/F1)에 대한 멜라닌 생성 저해시험, collagenase 및 elastase 활성 저해시험을 실시하였다. 그 결과, 합성된 coenzyme Qn유도체들의 농도에 따른 멜라닌 생성억제, collagenase와 elastase의 활성 저해시험 모두에서 우수한 활성을 보여주었다. Coenzyme Qn화합물들 중 coenzyme $Q_1$과 coenzyme $Q_2$는 멜라닌 생성억제와 elastase 활성저해활성이 다른 coenzyme Qn유도체들보다 우수하게 나타났다. 하지만 collagenase 활성저해시험에서는 coenzyme Qn 화합물들이 80-85%의 collagenase 활성저해력을 나타내었다. 또한 coenzyme Qn들의 생리활성을 비교하면, collagenase 저해활성시험을 제외한 다른 효소활성에서는 isoprene unit의 n수가 적을수록 더 우수한 활성이 있음을 확인하였으며, collagenase의 경우 coenzyme Qn 화합물의 생리활성이 모두 유사함으로 isoprene unit의 n수보다 coenzyme Qn화합물의 benzoquinone에 의한 효과로 생각된다. 이와 같은 결과로, coenzyme $Q_1$과 coenzyme $Q_2$는 앞으로 새로운 기능성 화장품으로 사용될 수 있을 것으로 사료된다.

• 미생물학회지
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• 제40권2호
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• pp.111-114
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• 2004

단삼(Salvia miltiorrhiza) 추출물의 치주질환유발 세균에 대한 생육저지 활성 및 collagenase저해 활성을 측정하였다. 단삼 에탄올 추출물은 치주질환유발 세균에 대해 우수한 생육저지 활성을 나타내었다. 유기용매 분획물중, hexane에서 생육억제능이 가장 우수하였다. 치주질환유발 세균 C. curvus, C. rectus, E. corrodens, F. nucleatum, P. gingivalis, P. intermedia 및 W. succinogenes에 대한 단삼 추출물의 최소생육저해농도(MIC)는 200,50,50,250,150,250및 200 ${\mu}g$/ml로 각각 우수한 생육억제능을 나타내었다. 단삼 유기용매 분획물의 collagenase저해 활성은 동일 농도의 minocycline에 비해 우수한 저해 활성을 나타내었으며, chloroform분획물의 경우 88.2%의 저해 활성을 나타내었다.

• Biomolecules & Therapeutics
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• 제14권1호
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• pp.36-39
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• 2006

Some flavones/flavonols were previously found to inhibit collagenase. To establish a therapeutic potential for skin inflammation, twenty-three synthetic flavone derivatives were examined for their inhibitory potential against collagenase from Clostridium histolyticum. From the results, it was found that most of them having various hydroxyl, methoxyl, methylsulfuryl and/or chloro substitution(s) on A- and B-rings were not efficient collagenase inhibitors. Among the synthetic flavones tested, only two synthetic derivatives, 3',4'-dihydroxyflavone and 5-hydroxy-4'-methoxyflavone, weakly inhibited bacterial collagenase (13-29% inhibition at 50-100 ${\mu}M$).

• Journal of Periodontal and Implant Science
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• 제29권1호
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• pp.31-40
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• 1999

Many factors may affect periodontal changes during the physiologic conditions of woman(e.g. puberty, menstrual cycle, pregnancy, menopause). Recently many research has focused on the immunological changes of host, but the exact mechanism is not clear. Collagen is a major constituent of periodontium, and collagenase specifically digests the collagen and plays a role in destruction of periodontal tissue. So, I suppose that it participates with the cytokines in the inflammation of gingiva and vascular response during the changes of female sex hormones. Because there are some evidences of the existence of the receptors of estrogen and progesterone in the gingiva, it may be a target tissue of female sex hormones. In this experiment, gingival fibroblast and periodontal ligament cell were cultured in the presence of various concentrations of estrogen or progesterone corresponding to the menstrual cycle and pregnancy. Collagenase activity of the supernatant of culture media was determined by Spectrophotometric collagenase assay. The enzyme activity was calculated by the % decrease of the coated collagen. 1. The estrogen at both concentrations had no effect on the activity of collagenase of the gingival fibroblast. 2. The progesterone had some effect on the collagenase activity of the gingival fibroblast at low and high concentration of menstrual cycle, and elevated the enzyme activity at all range of pregnancy concentrations. 3. In periodontal ligament cells, estrogen elevated the enzyme activity at the early pregnancy concentration and progesterone elevated at the concentration just before menstruation. In this experiment, pregesterone elevated the collagenase activity of gingival fibroblast and periodontal ligament cells. But the mechanism of the up-regulation of the enzyme activity was not confirmed. The more experiments of direct effect of progesterone on gingival at the molecular level(e.g. northern blot analysis) can reveal the exact mechanism.