• ALGAE
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• 제21권1호
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• pp.141-148
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• 2006

Benthic diatoms are known as a good food for shellfish in nature and in commercial hatchery of Jeju Island, Korea. Grammatophora marina is commonly found as dominant benthic micro-algae in Jeju coastal waters throughout the year. To know the best growth conditions of this species, culture was done in terms of three parameters; water temperature, salinity and nutrients. Each parameter was controlled by temperature of 15, 20 and 25°C; salinity of 25, 30 and 35 psu; and nutrient concentrations of 50, 100 and 200%. F/2 media was used with artificial seawater for the culture, which was continued for two weeks with L:D cycle 12:12 by using fluorescent light. Maximum specific growth rate was recorded 1.68 d–1 at temperature of 25°C with salinity of 35 psu and nutrient concentration of 200% on 6th day during the culture period. Maximum biomass was also observed 4.9 × 105 cells mL–1 in the same condition. This species may belong to the euryhaline and eutrophic habitat with warm condition. For nutritional aspects of this species, protein, lipid and carbohydrate were measured. The value of protein, lipid and carbohydrate was 4.96%, 15.82% and 5.65%, respectively. The antioxidant activities of 80% methanolic extract were 46.7%, 23.7% and 23.8% on DPPH (1,1-Diphenyl-2-picrylydrazy) radical, superoxide anion radical and hydrogen peroxide scavenging, respectively. Percentage metal chelating activity was 81.2%. Enzymatic extracts of Alcalase and Ultraflow showed remarkable scavenging activities on DPPH radical (86.5% and 57.2%, respectively), and superoxide anion scavenging activities were 45.3% and 41.4% from Kojizyme and Viscozyme extracts, respectively. Extract of Protomex revealed 24.8% activity on hydrogen peroxide and Neutase showed 30.8% on hydroxyl radical scavenging effects. Celluclast and Viscozyme extracts showed 33.2% and 32.1% activities on nitric oxide scavenging, respectively, while Alcalase showed 61.5% on metal chelating. This species contains higher lipids among the biochemical compounds and higher metal chelating activities from both 80% methanolic and enzymatic extracts.

• 한국해양생명과학회지
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• 제8권1호
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• pp.56-67
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• 2023

Marine macroalgae are important in coastal ecosystems and interact with marine microorganisms. In this study, we isolated fungi from seven types of marine macroalgae including Cladophora sp., Gloiopeltis furcate, Gracilariopsis chorda, Hydroclathrus clathratus, Prionitis crispata, Sargassum micracanthum, and Ulva lactuca collected in Korea. Morphological and phylogenetic analyses identified the isolates as four Aspergillus spp. (A. fumigatus, A. sydowii, A. tamarii, and A. terreus), three Penicillium spp. (P. crustosum, P. jejuense, and P. rubens), and Cladosporium tenuissimum. Among them, A. fumigatus TOP-U2, A. tamarii SH-Sw5, and A. terreus GJ-Gf2 strains showed the activities of all enzymes examined (amylase, chitinase, lipase, and protease). Based on the enzymatic index (EI) values in solid media, A. terreus GJ-Gf2 and C. tenuissimum UL-Pr1 exhibited the highest amylase and lipase activities, respectively. Chitinolytic activity was only observed in A. terreus GJ-Gf2, A. tamarii SH-Sw5, and A. fumigatus TOP-U2. Penicillium crustosum UL-Cl2 and C. tenuissimum UL-Pr1 showed the highest protease activities. To the best of our knowledge, this is the first report of lipolytic and proteolytic activities in a marine-derived C. tenuissimum strain. Overall, the fungal strains isolated from the marine macroalgae in this study actively produced industrially important enzymes.

• 미생물학회지
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• 제36권2호
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• pp.130-135
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• 2000

순천만 갯벌의 3개 지점의 표층(5cm)과 심층(20 cm)토양의 물리화학적 요인과 종속영양세균의 분포, 섬유소 분해율과 체외효소의 활성을 측정하였다. 온도, 수분함량, 인산염 인 그리고 유기물함량은 각각 -1~$30^{\circ}C$, 42.1~53.1%, 0.0779~0.1961mg/g, 그리고 1.99~7.64%로 나타나었다. 섬유소 films의 분해율은 7.7%~100%/month 범위로 1~2월에 최저 그리고 8~9월에 최고의 분해율을 나타냈다. 종속영양세균의 분포는 $0.87{\times}10^6 ~ 3.6{\times}10^7 $CUFs/g dry soil의 범위에 속해Tdmu 심층의 경우 표층에 비해 낮았다. MUF-기질의 분해율로 측정한 phoshatase, $\alpha$-D-gluocosidase, $\beta$-D-glucosidase, cellobiohydrolase 활성의 변화는 각각 152.23~1779.80 nMi/hr 2.67~202.18 nM/hr, 5.03~258.26 nM/hr, 3.42~63.07 nM/hr으로 모두 하계에 상대적으로 높은 값을 나타내었다. 섬유소 분해는 수온상승이 주된 요인이었으며, 정점간, 깊이간 큰 차이는 나타나지 않았다. 체외효소의 활성은 온도 및 섬유소 분해율과 높은 양의 상관관계를 나타냈다. 종속영양세균의 분포는 섬유소 분해와 체외효소의 활성과 높은 상괸은 나타나지 않았다.

• 한국미생물·생명공학회지
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• 제43권4호
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• pp.378-384
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• 2015

누룩은 전통주를 빚기 위한 발효제이다. 본 연구의 목적은 누룩 발효시 온도의 영향을 알아보기 위한 것이다. 누룩 발효는 $36^{\circ}C$에서 30일간 발효한 처리구(TN-A)와 $45^{\circ}C$에서 10일간 전발효 후, $36^{\circ}C$에서 20일간 후 발효시킨 처리구(TN-B)로 실험하였다. 누룩 발효시 온도에 따른 ${\alpha}$-amylase, glucoamylase, acidic protease 등 효소활성과 누룩의 대사산물을 측정하였다. 초기 효소활성은 온도에 상관없이 급격하게 증가하였고 발효 3일 후, TN-A 누룩의 효소활성은 일정하게 유지됐으나 TN-B 누룩은 발효기간이 끝나는 동안 점진적으로 이들 활성이 감소하였다. $^1H$-MNR을 이용한 대사체 분석결과, TN-A 누룩은 glucose, glycerol, fructose, mannitol, lactose가 초기에 급격히 증가한 후 감소하였으나, TN-B 누룩은 초기에 대사체들이 감소한 후, 발효기간 동안 일정하게 유지되었다. Glycine, proline, serine 유리아미노산은 TN-B보다 TN-A 누룩에서 보다 높게 나타났다. 밀누룩 발효시 약 $36^{\circ}C$의 일정한 온도는 전분 및 단백질분해 효소의 높은 활성을 유지하는데 적절하였다.

• BMB Reports
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• 제47권7호
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• pp.376-381
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• 2014

Enzymatic oxidation of pyrogallol was efficiently transformed to an oxidative product, purpurogallin (PPG). Here, the anticoagulant activities of PPG were examined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), and the activities of thrombin and activated factor X (FXa). And, the effects of PPG on expression of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were evaluated in tumor necrosis factor (TNF)-${\alpha}$ activated human umbilical vein endothelial cells (HUVECs). Treatment with PPG resulted in prolonged aPTT and PT and inhibition of the activities of thrombin and FXa, as well as inhibited production of thrombin and FXa in HUVECs. In addition, PPG inhibited thrombin-catalyzed fibrin polymerization and platelet aggregation. PPG also elicited anticoagulant effects in mice. In addition, treatment with PPG resulted in significant reduction of the PAI-1 to t-PA ratio. Collectively, PPG possesses antithrombotic activities and offers a basis for development of a novel anticoagulant.

• Journal of Oral Medicine and Pain
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• 제33권1호
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• pp.1-8
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• 2008

타액에 존재하는 여러 항균물질은 상호작용을 통하여 부가적이거나 상승작용을 나타내고 때로는 저해작용을 나타내는 것으로 알려져 있다. 본 연구는 in vitro 상에서 lysozyme과 peroxidase 사이의 상호작용을 효소활성 측면에서 조사하고자 시행되었다. Lysozyme과 peroxidase의 상호작용은 hen egg-white lysozyme(HEWL)과 bovine lactoperoxidase(bLP)를 혼합하는 방법으로 조사되었고, peroxidase system이 lysozyme에 미치는 영향은 potassium thiocyanate와 hydrogen peroxide를 추가적으로 첨가하는 방법으로 조사되었다. Lysozyme 활성은 Micrococcus lysodeikticus 기질용액의 혼탁도 변화를 측정하는 방법으로 측정되었고, peroxidase 활성은 NbsSCN 법으로 측정되었다. Wilcoxon signed rank 법을 이용하여 lysozyme과 peroxidase 효소활성 변화를 대조군과 비교하였다. 생리적 농도범위에서 bLP는 HEWL의 효소활성을 증가시켰으며(P < 0.05), 그 효과는 bLP의 농도증가에 따라 영향을 받았다. 하지만 HEWL는 bLP의 효소활성에 영향을 주지 못하였다. Thiocyanate는 HEWL의 효소활성에 영향을 주지 못하였고, potassium thiocyanate와 hydrogen peroxide를 추가한 peroxidase system도 HEWL 효소활성의 추가적인 상승을 유도하지는 못하였으며, hydrogen peroxide의 농도변화도 영향을 미치지 못하였다. 본 연구의 결과는 타액 항균 물질을 포함하고 있는 구강건강용품이나 구강에서 lysozyme과 peroxidase의 상호작용을 이해하는데 필요한 중요한 정보를 제공해준다.

• 한국수산과학회지
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• 제51권2호
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• pp.127-134
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• 2018

Arteriosclerosis is the major cause of coronary artery and cerebrovascular disease, which are leading causes of death. Pro-inflammatory cytokines induce injury to vascular endothelial cells by increasing cell adhesion molecules, leading to vascular inflammation, a major risk factor for the development of arteriosclerosis. In the current study, we investigated the inhibitory effect of enzymatic hydrolysate from Japanese mud shrimp Upogebia major on the inflammation of tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$)-stimulated human umbilical vein endothelial cells (HUVECs). We first evaluated the antioxidant and angiotensin I-converting enzyme (ACE) inhibitory activities of eight U. major enzymatic hydrolysates: alcalase, papain, ${\alpha}$-chymotrypsin (${\alpha}-Chy$), trypsin, pepsin, neutrase, protamex and flavourzyme. Of these, ${\alpha}-Chy$ exhibited potent antioxidant and ACE inhibitory activities. The ${\alpha}-Chy$ hydrolysate was fractionated by two ultrafiltration membranes of 3 and 10 kDa. The ${\alpha}-Chy$ hydrolysate of U. major and its molecular weight cut-off fractions resulted in a significant reduction in NO production and a decrease in cell adhesion molecules [vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and endothelial-selectin (E-selectin)] and pro-inflammatory cytokines [interleukin-6 (IL-6), interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1)] in $TNF-{\alpha}$-stimulated HUVECs. These results suggest that enzymatic hydrolysate from U. major can be used in the control and prevention of vascular inflammation and arteriosclerosis.

• 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.

• 한국해양바이오학회지
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• 제10권2호
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• pp.62-72
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• 2018

An active ingredient with hyaluronidase (HAse) inhibitory effect is one of the anti-aging approaches in cosmeceuticals. Here, red sea cucumbers (RSCs), Stichopus japonicus, from Jeju Island were evaluated to examine their HAse inhibitory and antioxidant activity effect. In this study, RSCs were extracted by six enzymatic hydrolysis (Alcalase; Al, Trypsin; Try, Neutrase; Neu, Pepsin; Pep, Alpha-chymotrypsin; Chy and Protamex; Pro). Alcalase hydrolysate (AlH) showed the highest antioxidant capacities for both of oxygen radical absorbance capacity (ORAC) and trolox equivalent antioxidant capacity (TEAC) methods, compared to those of other hydrolysates, at $66.59{\pm}0.78{\mu}M\;TE/mg$ and $135.78{\pm}3.24{\mu}M\;TE/mg$, respectively. Furthermore, AlH performed the highest capacity of HAse inhibitory with $IC_{50}$ value of 3.21 mg/ml. Thus, RSCs hydrolyzed by Al were chosen to determine the cellular antioxidant activity and hyaluronic acid (HA) production effect on Human immortalized keratinocyte cell line (HaCaT). The results showed that AlH improved the cell viabilities and intracellular reactive oxygen species (ROS) induced by 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH) were significantly decreased. In addition, AlH increased HA amount by regulating HYAL2 and HAS2 expressions in the HaCaT cells. Taken together, AlH of RSCs collected from Jeju Island showed HAse inhibitory and antioxidant activities against skin-aging which shows its potentials can be an optional natural bioactive ingredient for novel cosmeceuticals.

• 한국해양바이오학회지
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• 제7권1호
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• pp.19-27
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• 2015

The objective of the current study was to evaluate the inhibitory and antioxidant activities of powdered katsuobushi (dried bonito) protein hydrolysates and their corresponding fractions. The powdered katsuobushi (dried bonito) hydrolysates were obtained by enzymatic hydrolysis using Alcalase, ${\alpha}$-chymotrypsin, Neutrase, pepsin, papain, and trypsin. The antioxidant efficacy of the respective hydrolysates were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, superoxide, and alkyl radical-scavenging activities. Among the hydrolysates, the peptic-derived hydrolysate exhibited the highest antioxidant activity compared to other enzymatic hydrolysates. Therefore, the peptic-derived hydrolysate was further analyzed, and was found to contain an active peptide with an amino acid sequence identified as Pro-Met-Pro-Leu-Asn-Ser-Cys (756 Da). The purified peptides from powdered katsuobushi (dried bonito) had an $EC_{50}$ value of $105.82{\mu}M$, and exhibited an inhibitory effect against DNA oxidation induced by hydroxyl radicals. Taken together, these results suggests that powdered katsuobushi (dried bonito) could be used as a natural antioxidant in functional foods and prevent oxidation reactions in food processing.