• Title/Summary/Keyword: bioactivity

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Comparison of Enzymatic Activity and Cleavage Characteristics of Trypsin Immobilized by Covalent Conjugation and Affinity Interaction (공유결합과 친화력결합에 의한 고정화 Trypsin의 효소역가와 절단특성 비교)

  • Jang, Dae-Ho;Seong, Gi-Hun;Lee, Eun-Kyu
    • KSBB Journal
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    • v.21 no.4
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    • pp.279-285
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    • 2006
  • We investigated the effects of immobilization chemistry on the yield of immobilization and the bioactivity of the immobilized enzymes. Trypsin as a model protein and macroporous polymer beads(Toyopearl AF 650M, Tosho Co., Japan) was used as a model matrix. Four methods were used to immobilize trypsin; covalent conjugation by reductive amination(at pH 10.0 and pH 4.0) and affinity interaction via streptavidin-biotin, and double-affinity interaction via biotin-streptavidin-biotin system. The covalent conjugation immobilized $3{\sim}4$ mg/ml-gel, ca. 3-fold higher than the affinity method. However, the specific activity of the covalently(pH 10.0) and affinity-immobilized trypsin(via streptavidin-biotin) are ca. 37% and 50%, respectively, of that of the soluble enzyme(on the low-molecular-weight BAPNA substrate). When the molecular size of a substrate increased, the affinity-immobilized trypsin showed higher clavage activity on insulin and BSA. This result seemed to indicate the streptavidin-biotin system allowed more steric flexibility of the immobilized trypsin in its interaction with a substrate molecule. To confirm this, we studied the molecular flexibility of immobilized trypsin using quartz crystal microbalance-dissipation. Self-assembled monolayers were formed on the Q-sensor surface by aminoalkanethiols, and gultaraldehyde was attached to the SAMs. Trypsin was immobilized in two ways: reductive amination(at pH 10.0) and the streptavidin-biotin system. The dissipation shift of the affinity-immobilized trypsin was $0.8{\times}10^{-6}$, whereas that of the covalently attached enzyme was almost zero. This result confirmed that the streptavidin-biotin system allowed higher molecular flexibility. These results suggested that the bioactivity of the immobilized enzyme be strongly dependent on its molecular flexibility.

Controlled release of nerve growth factor from heparin-conjugated fibrin gel within the nerve growth factor-delivering implant

  • Lee, Jin-Yong;Kim, Soung-Min;Kim, Myung-Jin;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.40 no.1
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    • pp.3-10
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    • 2014
  • Objectives: Although nerve growth factor (NGF) could promote the functional regeneration of an injured peripheral nerve, it is very difficult for NGF to sustain the therapeutic dose in the defect due to its short half-life. In this study, we loaded the NGF-bound heparin-conjugated fibrin (HCF) gel in the NGF-delivering implants and analyzed the time-dependent release of NGF and its bioactivity to evaluate the clinical effectiveness. Materials and Methods: NGF solution was made of 1.0 mg of NGF and 1.0 mL of phosphate buffered saline (PBS). Experimental group A consisted of three implants, in which $0.25{\mu}L$ of NGF solution, $0.75{\mu}L$ of HCF, $1.0{\mu}L$ of fibrinogen and $2.0{\mu}L$ of thrombin was injected via apex hole with micropipette and gelated, were put into the centrifuge tube. Three implants of experimental group B were prepared with the mixture of $0.5{\mu}L$ of NGF solution, $0.5{\mu}L$ HCF, $1.0{\mu}L$ of fibrinogen and $2.0{\mu}L$ of thrombin. These six centrifuge tubes were filled with 1.0 mL of PBS and stirred in the water-filled beaker at 50 rpm. At 1, 3, 5, 7, 10, and 14 days, 1.0 mL of solution in each tubes was collected and preserved at $-20^{\circ}C$ with adding same amount of fresh PBS. Enzyme-linked immunosorbent assay (ELISA) was done to determine in vitro release profile of NGF and its bioactivity was evaluated with neural differentiation of pheochromocytoma (PC12) cells. Results: The average concentration of released NGF in the group A and B increased for the first 5 days and then gradually decreased. Almost all of NGF was released during 10 days. Released NGF from two groups could promote neural differentiation and neurite outgrowth of PC12 cells and these bioactivity was maintained over 14 days. Conclusion: Controlled release system using NGF-HCF gel via NGF-delivering implant could be an another vehicle of delivering NGF to promote the nerve regeneration of dental implant related nerve damage.

Quality characteristics and bioactivity analysis of habanero red pepper (Capsicum chinense Jacq.) through acetic acid bacteria fermentation (하바네로 고추 초산발효물의 품질 특성 및 생리활성 분석)

  • Park, Seul-Ki;Jo, Seung-Wha;Yim, Eun-Jung;Kang, Hyeon-Jin;Choi, Dong-Seong;Jeong, Do-Youn
    • Korean Journal of Food Science and Technology
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    • v.53 no.6
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    • pp.768-774
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    • 2021
  • Habanero red pepper (HRP)-fermented liquid with Acetobacter pasteurianus SRCM101474 was prepared, the quality characteristics measured and bioactivity analysis were measured. As the fermentation period increased, the number of viable cells increased, pH decreased, and acidity increased. After fermentation, the organic acid content of HRP was confirmed to be acetic acid with the highest content of 33413.54 ppm. Analysis of capsaicin and dihydrocapsaicin showed that the increase was significantly raised as acetic acid fermentation progressed. The total phenol and flavonoid contents as well as the antioxidant activity increased as the fermentation period passed. In addition, α-glucosidase inhibitory and pancreatic lipase inhibitory activities were significantly increased after fermentation. It was confirmed that it can be used as a functional material by measuring the physiological activities of antioxidant, anti-obesity, and anti-diabetes through fermentation, and the possibility of development as a spicy material and application product using it as capsaicin increases after fermentation.

Effect of Addition of β-TCP on Bioactivity and Mechanical Properties of Biodegradable PLA/β-TCP Composites (β-TCP 첨가량에 따른 생분해성 PLA/β-TCP 복합체의 생체활성과 기계적 특성 평가)

  • Moon, Hee-Jung;Kim, Seol-Ak;Kim, Dong-Ae
    • The Journal of the Korea Contents Association
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    • v.21 no.3
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    • pp.609-615
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    • 2021
  • The aim of the present study was to investigate the mechanical properties of melt-injected poly lactic acid (PLA) composites with β-tricalcium phosphate (β-TCP). The PLA mixed with calcined PLA/β-TCP powder to be contents of 0, 10, 30, 50 wt%, respectively, was dissolved in chloroform solvent under stirring for 24 h. Then the liquid mixtures were dropped into ethanol to extract solvent. After drying, the well-dispersed PLA/β-TCP composites were granulated and melt-injected to prepare specimens for various mechanical testing. PLA/β-TCP induced the precipitation of an apatite bone-mineral phase on the surface after immersion in a human simulated body fluid (SBF) for 90 days, showing in bioactivity. Mean various mechanical properties PLA/β-TCP composite were increased up to 10-30 wt% with significantly in part and composite were decreased 50 wt% of showing in mechanical properties. In conclusion, Over 30 wt% addition of β-TCP to PLA may be not advisable to improve the mechanical properties of melt-injected polymeric composites. Results indicated that β-TCP can be used considered as potential reinforcing agent for increasing mechanical properties for PLA. Therefore, it was suggest that the additional effects of β-TCP and research on a wide range of substances.

Food Functionality and In Vitro Bioactivity of Olive Flounder Paralichthys olivaceus Roe Concentrates Prepared by Cook-dried Process (가열-건조처리 넙치(Paralichthys olivaceus) 알 농축물의 식품기능성 및 생리활성)

  • In Seong Yoon;Sang in Kang;Jin-Soo Kim;In Sang Kwon;Hyeung Jun Kim;Min Soo Heu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.1
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    • pp.7-20
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    • 2023
  • Boil-dried concentrate (BDC) and steam-dried concentrate (SDC) were prepared from olive flounder Paralichthys olivaceus roe using the cook-dried process, and their food functionality and in vitro bioactivity were examined. The buffer capacity of BDC and SDC was found to be stronger in the alkaline region than in the acidic region, and the buffer capacity of SDC was superior to that of BDC. The water holding capacities of these concentrates were 7.6 and 7.4 g/g protein, respectively, both of which were significantly lower than that of freeze-dried concentrate (FDC). The solubility of BDC (13.4%) and SDC (12.7%), foaming capacity of BDC (107.7%) and SDC (110.6%), and oil-in-water emulsifying activity index of BDC (7.7 m2/g) and SDC (9.7 m2/g) were all significantly lower than the corresponding values for FDC (P<0.05). The lower food functionality of BDC and SDC compared with FDC can be attributed to the high-temperature denaturation of proteins during the cook-dried process. The 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging activities (IC50) of SDC (2.5 mg protein/mL) was 60.4 ㎍/mL, and the angiotensin I converting enzyme inhibitory activity was 80.9%. Olive flounder roe concentrates have good antioxidant and antihypertensive activities, and can be used as materials or ingredients in the processing of seafood and other foods to enhance protein contents and food functionality.

Cellular Interaction of In Situ Chitosan- and Hyaluronic Acid-Based Hydrogel

  • Noh, In-Sup
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.183-183
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    • 2006
  • Hyaluronic acid and chitosan-based poly(ethylene oxide) (HA-PEO and Chitosan-PEO) hydrogels have been employed as unique biomedical polymeric materials with properties such as bioactivity from polysaccharide, biocompatibility of HA and chitosan as well as PEO and control release of bioactive molecules from the hydrogel itself. We here examine in situ hydrogels based on hyaluronic acid and chitosan in terms of their synthesis, mechanical properties, morphologies and in vitro cellular interactions on their surface and inside. In vivo bone regeneration of HA-PEO and Chitosan-PEO hydrogels was compared with in mouse model.

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