• Title/Summary/Keyword: collagenase

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Purification and Characterization of a Collagenase from the Mackerel, Scomber japonicus

  • Park, Pyo-Jam;Lee, Sang-Hoon;Byun, Hee-Guk;Kim, Soo-Hyun;Kim, Se-Kwon
    • BMB Reports
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    • 제35권6호
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    • pp.576-582
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    • 2002
  • Collagenase from the internal organs of a mackerel was purified using acetone precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, gel filtration chromatography on a Sephadex G-100, ion-exchange chromatography on DEAE-Sephacel, and gel filtration chromatography on a Sephadex G-75 column. The molecular mass of the purified enzyme was estimated to be 14.8 kDa by gel filtration and SDS-PAGE. The purification and yield were 39.5-fold and 0.1% when compared to those in the starting-crude extract. The optimum pH and temperature for the enzyme activity were around pH 7.5 and $55^{\circ}C$, respectively. The $K_m$ and $V_{max}$ of the enzyme for collagen Type I were approximately 1.1 mM and 2,343 U, respectively. The purified enzyme was strongly inhibited by $Hg^{2+}$, $Zn^{2+}$, PMSF, TLCK, and the soybean-trypsin inhibitor.

유채박 추출물의 피부미용 효과에 관한 연구 (A Study on Skin Care Effects of Rapeseed Meal Extract)

  • 김선미;나명순
    • KSBB Journal
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    • 제28권3호
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    • pp.177-184
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    • 2013
  • This research was to investigate the inhibitory effects of tyrosinase, elastase and collagenase using rapeseed meal extract for the functional cosmetics. Gamma-tocopherol and alpha-tocopherol contents were 304.9 and 212.2 mg/kg, respectively. In the case of delta-tocopherol and plastochromanol-8, they were 12.1 and 35.7 mg/kg, respectively. The total phenol content of methanol extract was the highest (49.6 mg/g) which was about 4.96 fold higher than that of water extract. The maximum nitrite scavenging activities of methanol and acetone extract at pH 1.2 were 85.2 and 80.1%, respectively, at 8.0 mg/mL. When the extract concentration of rapeseed meal increased upto 2.0 mg/mL, cell viabilities did not appear to have any significant cytotoxic effect, irrespective of extracts. Tyrosinase and elastase inhibition activities increased from 25.2 to 42.5% and 25.3 to 48.0%, respectively, as methanol extract concentration increased from 0.5 to 1.0 mg/mL. The collagenase inhibition activities of methanol and acetone extracts at 1.0 mg/mL were 67.2 and 68.0%, respectively. These results showed that the methanol and acetone extract of rapeseed meal can be used as a new source of functional cosmetic agent.

Ginseng Intestinal Bacterial Metabolite IH901 as a New Anti-Metastatic Agent

  • Hideo Hasegawa;Sung, Jong-Hwan;Huh, Jae-Doo
    • Archives of Pharmacal Research
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    • 제20권6호
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    • pp.539-544
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    • 1997
  • Anti-metastatic activities of IH901, an intestinal bacterial metabolic derivative formed from Ginseng protopanaxadiol saponins, was determined in vitro and in vivo. Under in vitro conditions, IH901 inhibited the migration of bovine aortic endothelial cells 25 times stronger than suramin and suppressed the invasion of HT1080 human fibrosarcoma cells into reconstituted basement membrane components of Matrigel 1000 times stronger than RGDS peptide. IH901 also showed inhibitory effect on type-IV collagenase secretion from HT 1080 cells and platelet aggregation. When the anti-metastatic activity of IH901 was evaluated in comparison with that of 5-FU using a spontaneous lung metastatic model of Lewis lung carcinoma, the administration of IH901 (10 mg/kg p. o.) to tumor-bearing mice led to a significant decrease in lung metastasis (43% of untreated control), which was slightly more effective than that obtained with 5-FU (56% of control). Thus, IH901 seems to exhibit its anti-metastatic activity partly through the inhibition of tumor invasion which results from the blockade of type IV collagenase secretion and also through anti-platelet and anti-angiogenic activities.

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초임계 유체로 추출된 산삼 부정 배양근 오일의 효모균 처리에 의한 비사포닌계 지방산 함량 변화 및 Collagenase 및 Elastase 저해 활성 증대 (Changes in Non-saponin Fatty Acid Content and Increases in Inhibitory Activities of Collagenase and Elastase by Treatment with Saccharomyces cerevisiae of the Supercritical Fluid Extracted Oil of the Adventitious Roots Culture of Wild Mountain Ginseng)

  • 김철중;심재권;권경철;임정대;최선강;유창연;이재근
    • 한국약용작물학회지
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    • 제26권2호
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    • pp.170-180
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    • 2018
  • Background: To obtain useful cosmetic resources, this study aimed to determine the non-saponin fatty acid and inhibitory activities of collagenase and elastase by treatment of Saccharomyces cerevisiae in supercritical fluid extracted oil of the adventitious root culture of wild mountain ginseng. Methods and Results: We performed supercritical fluid extraction at various conditions such as pressure, temperature, time, and use of co-solvents, unlike the n-hexane extraction for the adventitious roots culture of wild mountain ginseng. The non-saponin-fatty acid obtained from the oil of the adventitious roots culture was incresed by treatment with S. cerevisiae. The supercritical fluid extraction was conducted using gas chromatography. Non-saponin-fatty acid content, in the oil of adventitious roots culture of wild mountain ginseng treated with S. cerevisiae for 2 days were three times higher than that in the control. In addition, the oil of the adventitious roots culture treated with S. cerevisiae was investigated for the anti-wrinkle effect by using collagenase and elastase. The oil of adventitious roots culture treated with S. cerevisiae exhibited higher collagenase and elastase inhibitory activities than those in the control. Conclusions: Supercritical fluid extracted oil of the adventitious roots culture of wild mountain ginseng treated with S. cerevisiae was found to have decreased ratio of saturated fatty acids and incresed ratio and content of unsaturated fatty acids increased. Furthermore, it showed anti-wrinkle effects in vitro.

헛개나무 추출물의 화장품 생리활성에 관한 연구 (Study of Cosmeceutical Activities of Hovenia dulcis var. koreana Nakai Extracts)

  • 김세현;전동하;장민정;이진태;이창언;한진규;김진철;이도형
    • 한국산림과학회지
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    • 제99권6호
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    • pp.836-842
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    • 2010
  • 본 연구는 기능성 식 약용 소재로 널리 이용되는 헛개나무의 화장품원료로의 개발을 위하여 다양한 생리활성을 측정하였다. 헛개나무 과병과 뿌리의 열수추출물 및 에탄올추출물에 대한 DPPH법, ABTS법을 이용하여 항산화활성 측정시 열매, 뿌리 모두 열수추출물에서 뛰어난 항산화효과를 나타냈으며, 멜라닌 색소합성에 관여하는 tyrosinase의 활성을 측정한 결과 tyrosinase 저해활성은 과병에탄올추출물(HDFE) 500 mg/L에서 32.0%의 억제능을 확인 할 수 있었고, 뿌리에탄올추출물(HDRE)은 21.8%의 억제능을 보였다. 더 높은 농도인 1,000 mg/L에서는 과병열수추출물(HDFW), HDFE 순으로 저해활성이 나타났다. 주름개선효과에 관한 활성은 collagenase와 elastase 저해활성을 통해 측정하였다. 그 결과 elastase에서 과병에탄올 추출물을 제외한 모든 추출물이 두 효소를 모두 저해하는 것으로 나타났으며, 같은 농도에서는 collagenase의 활성이 더 크게 저해됨을 보였다. 이와 같은 결과를 통해 헛개나무추출물의 항산화활성과 미백 및 주름개선효과를 이용하여 기능성 화장품으로 활용이 가능할 것으로 사료된다.

왕겨초액의 대량생산과 항노화 및 미백 효과 (Mass Production of Chaff-vinegar and Its Effect of Anti-Aging and Whitening)

  • 황재규;윤종국;김숙경;이상한;한길환
    • 한국미생물·생명공학회지
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    • 제40권3호
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    • pp.208-214
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    • 2012
  • 본 연구는 왕겨를 이용한 항산화, 항노화에 효과적인 초액을 대량생산하여 기능성을 검증하고 왕겨초액 내의 기능성물질을 분석하여 사업화의 가능성을 타진하고자 하였다. 정제된 왕겨초액을 분획하여 기능성이 확인된 부분을 HPLC 기기를 이용하여 확인한 결과 caffeic acid 외에 7종의 polyphenol 유래물질에서 기능성을 확인하였다. 기능성 화장품소재 시험으로는 세포 생존율 측정(MTT assay), 주름개선 (collagenase inhibition assay, elastase inhibition assay), 미백(tyrosinase inhibition assay), 항산화 시험(DPPH free radical scavenging assay, SOD-like activity, xanthine oxidase inhibition assay)이 실시하였다. 피부주름형성은 콜라겐의 합성과 분해가 중요한 원인으로 작용한다고 알려져 있지만, 최근 많은 연구에서 엘라스틴의 재형성과 분해 또한 주름형성 기전에서 중요한 작용을 하는 것으로 보고되고 있다. 왕겨초액은 $100{\mu}L/mL$의 농도에서 collagenase 저해효과가 100%, elastase 저해효과가 55% 이상으로 주름개선 효과가 우수하였다. 왕겨초액은 $100{\mu}L/mL$의 농도에서 tyrosine에 대한 tyrosinase 저해효과가 80% 이상으로 미백효과가 우수하게 나타났다. DPPH radical 소거율, xanthine oxidase 저해와 SOD-like 활성을 이용하여 항산화 효과를 측정하였다. 왕겨초액 $100{\mu}L/mL$의 농도에서 DPPH radical 소거율이 80%, xanthine oxidase 저해효과가 80%, 그리고 SOD-like 활성이 100%를 나타내었다. 따라서 본 연구를 통하여 정제 왕겨초액을 이용한 산업화를 위한 가능성을 타진하여 효과가 좋음을 확인할 수 있었다. 이와 같은 결과로 왕겨초액을 이용한 기능성 화장품의 소재로 뿐만 아니라, 식품 및 의약외품의 품목허가를 통하여 고부가가치의 상품화소재로의 가능성이 있다고 사료됩니다.

Establishment and Identification of a Debao Pony Ear Marginal Tissue Fibroblast Cell Line

  • Zhou, X.M.;Ma, Y.H.;Guan, W.J.;Zhao, D.M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권10호
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    • pp.1338-1343
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    • 2004
  • The Debao pony ear marginal tissue fibroblast cell line (NDPEM 2/2) was uccessfully established using either primary explant technique or collagenase technique. The characterizations of the cell line were identified as following: the cells were adherent and of density limitation; population doubling time (PDT) of cells made with the two techniques were 35.9 h and 48 h, respectively; chromosome analysis showed that the frequency of cell chromosome number to be 2n=64 was 91.3%-92.8%. Confirmed by isoenzyme analysis, this cell line had no cross- contamination. Tests for microbial contamination from bacteria, fungi, virus or mycoplasma were negative. This newly established cell line meets all the standard quality controls of ATCC. It will provide a precious genetic resource for the conservation of the Debao pony breed, as well as effective experimental material for genetic studies on Debao ponies.

가지 열수추출물의 피부생리활성에 관한 연구 (A Study of Physiological Activities of the Thermal Treated Eggplant on the Skin)

  • 김란
    • KSBB Journal
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    • 제31권3호
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    • pp.151-157
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    • 2016
  • The purpose of the present research was to investigate the physiological activities of the thermal treated eggplant on the skin. Five minute of thermal treatment at $100^{\circ}C$ had the highest polyphenol content of eggplant. However, below and over 5 min of thermal treatment time, they did not increase. When water and ethanol extracts were used, the maximum DPPH radical scavenging activities were obtained, 66.3 and 62.8%, respectively. Among various extracts, the acetone extract gave the highest cosmetic activity. Especially, when acetone extract (15.0 mg/mL) was used, the maximum inhibition activities of tyrosinase, elastase, and collagenase were obtained, 83.4, 78.2 and 62.5%, respectively. These results suggest the anti-wrinkle and whitening and effects of acetone extract were excellent. Therefore, it is should be considered as a promising candidate for novel cosmetic agents.