• Title/Summary/Keyword: enzymatic activities

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Growth Characteristics, Bio-chemical Composition and Antioxidant Activities of Benthic Diatom Grammatophora marina from Jeju Coast, Korea

  • Affan, Abu;Karawita, Rohan;Jeon, Yu-Jin;Kim, Bo-Young;Lee, Joon-Baek
    • ALGAE
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    • v.21 no.1
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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.

Investigation of Enzymatic Activities in Marine Algae-Derived Fungi

  • Dawoon Chung;Woon-Jong Yu;Hyeong Seok Jang;Yong-Min Kwon;Seung Seob Bae;Grace Choi
    • Journal of Marine Life Science
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    • v.8 no.1
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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.

Cellulose Degradation and Extracellulat Enzymatic Activity of the Mud Flat in Sunchon Bay (순천만 갯벌 토양의 섬유소 분해능 및 체외효소 활성)

  • 백근식;최지혁;성치남
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.130-135
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    • 2000
  • Decomposition rate of organic matiter in the mud flat of Sunchon Bay was estimated. Physicochemical parameters, cellulose degradation rate. distribution of heterotrophic bacteria, and extracellular enzymatic activities were measured from August 1997 to July 1998. Soil temperatures, water contents, concentration of $PO_4$-P and organic matter were -1-~$30^{\circ}C$, 42.1-53.1%, 0.0779-0.1961 mgig and 1.99-7.64%, respectively. Decomposition rate of cellulose film ranged from 7.7 to 100%imonth, high in summer and low in winter. The number of heterotrophic bacteria ranged from $0.87{\times}10^6 to 3.6{\times}10^7 $CUFsIg dq soil. Enzymatic activities of phosphatase, $\alpha$-D-gluEosidase, $\beta$-D-glucosidase and cellobiohydrolase, which were measured as decomposition rate of methylumbelliferyl(MLiF)-substrate, were 152.23-1779.80 nMIhr, 2.67-202.18 nM/hr, 5.03-258.26 M h r and 3.42-63.07 nM/hr, respectively Cellulose degradaaon rate and extracellular extracellular enzymatic activities were conelated with each other, and showed high correlation coefticiency with soil temperature.

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Effects of Temperature on the Changes of Enzymatic Activities and Metabolite during Wheat nuruk Fermentation (밀누룩 발효기간 동안 효소와 대사체 변화에 대한 온도의 영향)

  • Lee, Se Hee;Baek, Seong Yeol;Kang, Ji-Eun;Jeon, Che Ok;Kim, Dae-Hyuk;Kim, Myoung-Dong;Yeo, Soo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.378-384
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    • 2015
  • Nuruk is a fermentation agent, which has been used for the production of traditional Korean alcoholic beverages. The objective of this study was to investigate the effects of temperature on nuruk fermentation. One wheat nuruk sample was fermented at $36^{\circ}C$ for 30 days (TN-A) and another at $45^{\circ}C$ for 10 days followed by $36^{\circ}C$ for 20 days (TN-B). The activities of ${\alpha}$-amylase, glucoamylase, and acidic protease, as well as metabolite contents were measured. Initially, the enzymatic activities increased rapidly regardless of the fermentation temperature. After 3 days of fermentation, the enzymatic activities were maintained in TN-A, but gradually decreased in TN-B until the end of fermentation process. Metabolite analysis using $^1H$-NMR showed that the levels of glucose, glycerol, fructose, mannitol, and lactose initially increased quickly and then decreased in TN-A. However, they initially decreased and then were maintained over the fermentation period in TN-B. The contents of glycine, proline, and serine were higher in TN-A than in TN-B. This study suggests that a constant temperature of approximately $36^{\circ}C$ is appropriate for achieving high amylolytic and proteolytic activities in the production of wheat nuruk.

Antiplatelet and antithrombotic activities of purpurogallin in vitro and in vivo

  • Ku, Sae-Kwang;Bae, Jong-Sup
    • BMB Reports
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    • v.47 no.7
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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.

Influences of Peroxidase on Lysozyme Activity (Peroxidase가 Lysozyme 활성에 미치는 영향)

  • Lee, Sang-Goo;Kim, Hyung-Il;Kho, Hong-Seop
    • Journal of Oral Medicine and Pain
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    • v.33 no.1
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    • pp.1-8
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    • 2008
  • It is well known that many antimicrobial proteins in saliva interact with each other. The purpose of the present study was to investigate the interactions of lysozyme with peroxidase in the aspects of enzymatic activity in vitro. The interactions of lysozyme with peroxidase were examined by incubating hen egg-white lysozyme(HEWL) with bovine lactoperoxidase(bLP). The influence of peroxidase system on lysozyme was examined by subsequent addition of potassium thiocyanate and hydrogen peroxide. Lysozyme activity was determined by turbidity measurement of a Micrococcus lysodeikticus substrate suspension. Peroxidase activity was determined with an NbsSCN assay. The Wilcoxon signed rank test was used to analyze the changes of enzymatic activities compared with their controls. bLP at physiological concentrations enhanced the enzymatic activity of HEWL(P < 0.05) and its effect was dependent on the concentration of peroxidase. However, HEWL did not affect the enzymatic activity of bLP. Thiocyanate did not affect the enzymatic activity of HEWL, either. The addition of potassium thiocyanate and hydrogen peroxide did not lead to additional enhancement of the enzymatic activity of HEWL. The changes of hydrogen peroxide concentration in the peroxidase system did not affect the enzymatic activity of HEWL. Collectively, despite an in vitro nature of our study, the results of the present study provide valuable information on the interactions of lysozyme and peroxidase in the aspects of enzymatic activity in oral health care products and possibly in the oral cavity.

Inhibition Effect of Enzymatic Hydrolysate from Japanese Mud Shrimp Upogebia major on TNF-α-induced Vascular Inflammation in Human Umbilical Vein Endothelial Cells (HUVECs) (혈관내피세포에서 TNF-α로 유도되는 혈관염증에 대한 쏙(Upogebia major) 효소가수분해물의 억제 효과)

  • Kim, So-Yeon;Yang, Ji-Eun;Song, Jae-Hee;Maeng, Sang-Hyun;Lee, Ji-Hyun;Yoon, Na-Young
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.2
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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.

Biochemical and Genetic Characterization of Arazyme, an Extracellular Metalloprotease Produced from Serratia proteamaculans HY-3

  • Kwak, Jang-Yul;Lee, Ki-Eun;Shin, Dong-Ha;Maeng, Jin-Soo;Park, Doo-Sang;Oh, Hyun-Woo;Son, Kwang-Hee;Bae, Kyung-Sook;Park, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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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.

Hyaluronidase Inhibitory and Antioxidant Activities of Enzymatic Hydrolysate from Jeju Island Red Sea Cucumber (Stichopus japonicus) for Novel Anti-aging Cosmeceuticals

  • Ding, Yuling;Jiratchayamaethasakul, Chanipa;Kim, Eun-A;Kim, Junseong;Heo, Soo-Jin;Lee, Seung-Hong
    • Journal of Marine Bioscience and Biotechnology
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    • v.10 no.2
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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.

Characterization of an antioxidant peptide from katsuobushi (dried bonito) protein hydrolysates

  • Lee, Jung Kwon;Jeon, Joong-Kyun;Byun, Hee-Guk
    • Journal of Marine Bioscience and Biotechnology
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    • v.7 no.1
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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.