• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.5
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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.

• Journal of Life Science
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• v.15 no.4 s.71
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• pp.657-663
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• 2005

Collagenases are generally defined as enzymes that are capable of degrading the polypeptide backbone of native collagen under conditions that do not denature the protein. An extracellular collagenase-producing bacterial strain was isolated from kimchi and identified to be Bacillus subtilis JS-17 through morphological, cultural, biochemical characteristics and 16S rDNA sequence analysis. Optimum culture condition of Bacillus subtilis JS-17 for the production of collagenase was $1.5\%$ fructose, $1\%$ yeast extract, $0.5\%\;K_2HPO_4,\;0.4\%\;KH_2PO_4,\;0.01\%\;MgSO_4\cdot7H_2O,\;0.01\%\; MnSO_4\cdot4H_2O,\;,0.1\%$ citrate and $0.1\%\;CaCl_2$. The production of collagenase was optimal at $30^{\circ}C$ for 72 hr. A collagenase was isolated from the culture filtrate of Bacillus subtilis JS-17. The enzyme was purified using Amberlite IRA-900 column chromatography, Sephacryl S-300 HR column chromatography and DEAE-Sephadex A-50 column chromatography The purified collagenase has an specific activity 192.1 units/mg. The molecular weight of the purified enzyme was estimated to be 28 kDa by SDS-PACE. The purified collagenase has $100\%$ activity up to $55^{\circ}C$.

• Applied Biological Chemistry
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• v.51 no.3
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• pp.164-170
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• 2008

Coenzyme $Q_{10}$ and six derivatives of coenzyme Qn were synthesized and tested for their inhibitory effects on melanogenesis occurred in murine melanoma (B16/F1) cells and on collagenase/elastase activities as well. As the result, synthetic coenzyme Qn showed a potent inhibitory effect on melanin formation, collagenase and elastase activities in all tested concentrations. Among these synthetic compounds, coenzyme $Q_1$ and coenzyme $Q_2$ potentially inhibited melanin formation and elastase activity when compared to other coenzyme Qn derivatives. For the collagenase activities, all coenzyme Qn derivatives inhibited 80-85% of controls. As compared, coenzyme Qn derivatives exhibited strong inhibitory activities with the decrease of isoprenoid unit number of coenzyme Qn derivatives except for collagenase activity. For the inhibition of collagenase activity, moiety of benzoquinone might be considered as the active functional group. Taken together, coenzyme $Q_1$ and coenzyme $Q_2$ might be used for functional cosmetics.

• The Korean Journal of Physiology and Pharmacology
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• v.6 no.5
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• pp.269-274
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• 2002

A large number of factors such as osteotropic hormones, cytokines, or growth factors are related to the bone remodeling which is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. Recent investigations have indicated that cytokines such as $interleukin-1{\beta}\;(IL-1{\beta})$ and tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ play a potential role in the bone resorption associated with a variety of pathological conditions such as inflammatory osteolytic disease. Collagen is the most abundant protein of the extracellular matrix of bone, and the participation of collagenase in bone resorption has been widely investigated. In this study, effects of $IL-1{\beta}$ and $TNF-{\alpha}$ on the release of collagenase from osteoblastic cells were measured. The gelatinase activity was also measured by gel substrate analysis (zymography) after electrophoresis of conditioned media of osteoblastic cell culture. $IL-1{\beta}$ increased the collagenase activity in ROS17/2.8 and HOS cell culture. $TNF-{\alpha}$ also increased the collagenase activity of osteoblastic cells. When two kinds of cytokines were treated simultaneously in the culture of osteoblastic cells, synergistic increase of collagenase activity was seen in ROS17/2.8 cells. $IL-1{\beta}$ and $TNF-{\alpha}$ significantly increased the collagenase activity after 6 hour treatment in the osteoblastic cell culture, and there was no additional increase according to the culture period. Osteoblastic cells released the gelatinase and molecular weight of this enzyme was measured about 70 KDa as assessed by zymogram. $IL-1{\beta}$ and $TNF-{\alpha}$ showed increase of the gelatinase activity produced by ROS17/2.8 and HOS cells. Taken together, this study suggested that $IL-1{\beta}$ and $TNF-{\alpha}$ can modulate bone metabolism, at least in part, by increased release of collagenase and gelatinase from osteoblasts.

• Journal of fish pathology
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• v.12 no.2
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• pp.83-88
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• 1999

Green tea extract has been suggested to possess properties of collagenase inhibition and antimicrobial effect to fish pathogenic bacteria. Most fish pathogenic bacteria showed collagenase activities of 0.08-0.7 unit/ml. Among them, Edwardsiella tarda produced large amount of collagenase in the ST medium at $25^{\circ}C$ after 16 hrs. Biosynthesis of the enzyme from E. tarda was decreased to 1/3 by addition of 0.08-0.8 mg/ml of tea extract in the culture medium. The collagenase activity was inhibited almost 100% in the reaction mixture by 0.2 mg/ml of the extract. Then, the minimal inhibition concentration against E. tarda growth appeared as 8 mg/ml of the tea extract in the culture medium.

• The korean journal of orthodontics
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• v.28 no.1 s.66
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• pp.165-174
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• 1998

The turnover of collagen is controlled by the balance between collagen synthesis and degradation. The production of collagenase (matrix metalloproteinase-1) and its inhibitor, tissue inhibitor of matrix metallopmteinase-1 (TIMP-1) are one of the substances which regulate this balance. The periodontal ligament fibroblast plays an important role in collagen metabolism during orthodontic treatment and is believed to be an origin of the osteoblast in the alveolar bone. The collagenase secreted by the periodontal ligament fibroblast and the osteoblast initiates the bone resorption by removing the osteoid layer in the alveloar bone. The interleukin-$1{\beta}$ is secreted by the macrophage during orthodontic treatment. The present study was undertaken to assess the effect of mechanical stress and interleukin-$1{\beta}$ on the expression of collagenase and TIMP-1 in the periodontal ligament fibroblasts using reverse transcription polymerase chain reaction and immunohistochemical staining. The periodontal ligament fibroblasts were stitched by placing the $Petriperm dish^{\circledR}$ dish on the top of spheroidal convex watch glass ($5\%$ surface increase) and tented with interleukin-$1{\beta}$ (1.0 ng/ml), or treated with both of them. Treatment with mechanical stress and/or interleukin-$1{\beta}$ resulted in increased collagenase mRNA expression. The mechanical stress treated group (1.61, 1.62, 1.37 fold increase), the interleukin-$1{\beta}$, tented group (1.68, 1.60, 3.78 fold increase), the mechanical stress and interleukin-$1{\beta}$ treated group (1.89, 1.72, 5.48 fold increase) induced increases in collagenase mRNA compared with the control group after 2, 4, 8 hours respectively. But TIMP-1 mRNA expressions at experimental groups were decreased after 2, 4 hours and increased after 8 hours. The mechanical stress treated group (0.16, 0.49 fold decrease and 3.77 fold increase), the interleukin-$1{\beta}$ treated group (0.15,0.44 fold decrease and 4.46 fold increase), the mechanical stress and interleukin-$1{\beta}$ tented group (0.15, 0.69 fold decrease and 4.81 fold increase) induced changes in TIMP-1 mRNA compared with the control group after 2, 4, 8 hours, respectively. Immunohistochemical stain showed that increased collagenase and TIMP-1 staining of the mechanical stress tented group, the interleukin-$1{\beta}$ treated group, and the mechanical stress and interleukin-$1{\beta}$ treated group compared with that of the control group after 8 hours. These findings suggest that mechanical stress and interleukin-$1{\beta}$ regulate expression of collagenase and TIMP-1.

• The korean journal of orthodontics
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• v.20 no.2
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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.

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.245-252
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• 2006

A bacterial strain, identified as Staphylococcus aureus JJ-11, producing collagenase was isolated out of 40 persons having skin troubles. S. aureus JJ-11 produced collagenase optimally in the media containing 1.5%(w/v) gelatin, 1%(w/v) yeast extract, 0.4%(w/v) $K_2HPO_4$, 0.005%(w/v) $NiSO_4{\cdot}6H_2O$ at $37^{\circ}C$ for 18 hrs. The collagenase produced by Staphylococcus aureus JJ-11 was purified at 6.66-folds purity through application of chromatography with Amberlite IRA-900 and Sephacryl S-300 HR columns. The molecular weight of the partially purified enzyme was estimated to be 62 kDa by SDS-PAGE. The protein exhibited optimum enzymatic activity at pH 7.0, and showed a stable activity at pH 4-8. The optimum temperature for collagenase was at $37^{\circ}C$, and activity was maintained upto $40^{\circ}C$. The enzyme activity was slightly elevated in the presence of divalents such as, $Fe^{2+},\;Co^{2+}\;and\;Ba^{2+}$ However, the activity was inhibited in the presence of $Sr^{2+}\;or\;Hg^{2+}$. The inhibition of activity by O-phenanthroline and EDTA suggested that the enzyme may contain metal which is required for activity. The enzyme showed the highest activity when insoluble collagen (type I) was, used as a substrate.

• Advances in environmental research
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• v.11 no.1
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• pp.1-16
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• 2022

Amongst 27 isolates from deteriorated leather samples, Arthrobacter creatinolyticus KP015744 zzx28 was found to be an efficient collagenase producer. Collagenase production of 13.33 µmoles/min was shown at an optimum temperature at 37℃ after 72h and at pH 7.5 by using 2 ml/dL inoculum in 10 mg/ml collagen peptide type I as a substrate. In presence of Hg²⁺, EDTA and 𝛽-mercaptoethanol the collagenase production by the isolate was strongly inhibited however Fe²⁺, Ca²⁺and DMSO enhanced production of the enzyme. Specific activity was found to be 19.46×10³ U/mg and molecular weight 66 kD by SDS PAGE. Isolate also has potential to hydrolyze keratin which is another important protein found in leather. Experimental results propose that collagenase can be effectively used as a tool for collagen and keratin rich solid waste treatment.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.111-114
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• 2004

This study was carried out to evaluate the inhibition of growth and collagenase activity of Salvia miltiorrhiza Bunge against microorganisms causing periodontal diseases. The ethanol extract of Salvia miltiorrhiza showed sig-nificant growth inhibition against microorganisms causing periodontal diseases. Ethanol extract was further fractionated with organic solvents in the order of hexane, chloroform and ethyl acetate. Among the fractions tested, the hexane fraction showed the highest cell growth inhibition. The minimal inhibitory concentration (MIC) of the extract against C. curvus, C. rectus, E. corrodens, F nucleatum, P. gingivalis, P. intermedia and W. succinogenes were 200, 50, 50, 250, 150, 250 and 200 ${\mu}g$/ml, respectively. The inhibition of collagenase activity by organic solvent fractions were higher than that of minocycline, and the inhibition ratio of collagenase activity was $88.2{\pm}2.1$ % in the chloroform fraction.