• Title/Summary/Keyword: thrombin

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Evaluation of Anti-oxidant, Anti-microbial and Anti-thrombosis Activities of Fruit, Seed and Pomace of Schizandra chinensis Baillon (오미자 열매, 씨, 착즙 후 박의 항산화, 항균 및 항혈전 활성 평가)

  • Kim, Mi-Sun;Sung, Hwa-Jung;Park, Jong-Yi;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.27 no.2
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    • pp.131-138
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    • 2017
  • In this study, for the efficient use of the byproduct of the omija (Schizandra chinensis Baillon: SC) processing industry, the ethanol extracts of the fruit (F), seed (S), and pomace (P) of SC were prepared, and their useful bioactivities were evaluated. For F-SC, S-SC, and P-SC, the extraction yields were 28.3%, 22.1%, and 7.2%, respectively, and the polyphenol contents were 8.81, 37.22, and 9.20 mg/g, respectively. The total flavonoid content in P-SC (4.31 mg/g) was 3.5-fold higher than that in F-SC (0.76 mg/g). In an antioxidation activity assay, P-SC showed stronger radical scavenging activities against DPPH anion, ABTS cation, and nitrite and stronger reducing power activities than the other extracts. The calculated concentration required for 50% radical scavenging activity, $RC_{50}s$, of P-SC for DPPH anion, ABTS cation, and nitrite was 226.2, 192.5, and $92.5{\mu}g/ml$, respectively. In an antimicrobial activity assay, F-SC, S-SC, and P-SC showed similarly strong growth inhibitions against Bacillus subtilis and P. vulgaris at a concentration of 0.5 mg/disc. F-SC and P-SC showed 15-fold extended time in thrombin, prothrombin, and activated partial thromboplastin time assays at a concentration of 5 mg/ml. The anticoagulation activity of P-SC (2.5 mg/ml) was comparable to that of aspirin (1.5 mg/ml). Furthermore, F-SC and S-SC showed very good platelet aggregation inhibitory activities. F-SC, S-SC, and P-SC did not show significant hemolysis against human red blood cell up to a concentration of 0.5 mg/ml. These results suggest that S-SC and P-SC, both of which are byproducts of the omija processing industry, show strong potential as novel antioxidant, antimicrobial, and antithrombosis agents.

Changes in the in vitro Antioxidant and Antithrombosis Activities of Salicornia europaea According to Harvest Time (수확시기에 따른 함초의 항산화 및 항혈전 활성의 변화)

  • Kim, Mi-Sun;Lee, Jung-Nam;Seong, Ha-Jung;Kim, Deuk Hoi;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1033-1040
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    • 2016
  • In Korea, the aerial parts of the halophyte Salicornia europaea, known as hamcho, are used in salads in April–June and in oriental medicine in September–October In this study, with the aim of developing functional foods to aid blood circulation, hot water extract (HWE) and ethanol extract (EE) were prepared using hamcho harvested from the fields of Shinan, Jeonnam, Korea on 5th April (HWE-04, EE-04), 5th July (HWE-06, EE-06), 5th August (HWE-08, EE-08), 5th September (HWE-09, EE-09), and 5th October (HWE-10, EE-10), and their antioxidant and antithrombosis activities were evaluated. Among the HWEs, HWE-10 showed the highest concentration of total polyphenols and total flavonoids (22.4 and 17.6 mg/ml, respectively), and EE-09 had the highest concentration among the EEs (20.1 and 19.3 mg/ml, respectively). Among the HWEs and EEs, HWE-08 and EE-08 had the highest total sugar and reducing sugar content. In the antioxidation assay, HWE-10 and EE-09 showed strong reducing power, as well as DPPH, ABTS, and nitrite scavenging activities. The calculated RC50s of EE-09 against DPPH, ABTS, and nitrite were 578, 277, and 68.8 μg/ml, respectively. The antithrombosis activity assay revealed that HWE-04, HWE-06, EE-04, and EE-06 had anticoagulation activity against coagulation factors and that HWE-08, HWE-09, EE-08, and EE-09 expressed strong thrombin inhibitory activity, which was comparable to the antithrombosis activity of aspirin. In addition, EE-06 and HWE-08 exhibited strong aggregation inhibitory activities against human platelets. The results suggest that extract from hamcho harvested in particular periods and prepared using a defined solvent has strong potential as a novel food ingredient and an antioxidant and antithrombosis agent.

Anti-thrombotic Activities of Hot-water Extracts Prepared from Various Parts of Nelumbo nucifera Gaertner (다양한 부위의 연(Lotus) 열수 추출물의 항혈전 활성)

  • Ahn, Seon-Mi;Sung, Hwa-Jung;Kim, Jong-Sik;Park, Jong-Yi;Sohn, Ho-Yong
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1156-1162
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    • 2018
  • To investigate the anti-thrombotic activities of the lotus (Nelumbo nucifera Gaertner), various hot-water extracts were prepared from the leaf, pod of seed (PS), seed, embryo of seed (ES), root, and the node of root (NR) of the lotus. The highest extraction ratio was found in the NR (20.3%), followed by the seed, root, leaf, ES, and PS. These extracts had pH and acidity levels ranging from 5.6~6.5 and 0.06~0.20%, respectively. The seed extract showed 70% brix, whereas the leaf and PS extracts showed less than 0.1% brix. The highest contents of total polyphenol (179.7 mg/g), total flavonoids (161.4 mg/g), and reducing sugar (161.4 mg/g) were observed in the leaf extract, and the highest total sugar content (873.0 mg/g) in the seed extract. Anti-coagulation assay of the extracts of NR, leaf, and PS showed strong activities. In particular, at a concentration of 5 mg/ml, the PS extract had 15-fold extended thrombin, prothrombin, and activated partial thromboplastin times. However, only the ES extract showed activities inhibitory to platelet aggregation, with treatment with 0.25 mg/ml of ES extract decreasing platelet aggregation to 25.1%, a reduction comparable to that effected by aspirin. The extracts other than the seed extract showed no hemolysis activities against human RBC at treatments of up to 1 mg/ml. These results suggest that the NR, PS, seed, and ES, all byproducts of the lotus agriculture industry, have high potential as novel sources of anti-thrombotic agent.

A Comparison of the Components and Biological Activities in Raw and Boiled Red Beans (Phaseolus radiatus L.) (생팥과 증자팥의 성분 및 생리활성 비교)

  • Lee, Ryun Kyung;Kim, Mi-Sun;Lee, Ye-Seul;Lee, Man-Hyo;Lee, Jong Hwa;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.162-169
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    • 2014
  • In the course of study for the development of functional food using red beans (azuki beans, Phaseolus radiatus L.), the ethanol extracts from raw-red bean (RRB) and boiled-red bean (BRB) were prepared, and the components and various biological activities of both were compared. It was observed that the extraction yield, and the total polyphenol content, of the BRB were 1.2 times higher than that of the RRB. However, the contents of total flavonoid, total sugar and reducing sugar in the BRB were 30, 27.9 and 30.8% respectively when compared with those of RRB. In relation to antioxidative activity, both RRB and BRB exhibited moderate DPPH anion, ABTS cation, and nitrite scavenging activities and reducing power, though in all cases RRB demonstrated stronger activities than BRB. The extracts of RRB and BRB did not reveal any antimicrobial activities. In a ${\alpha}$-amylase inhibitory activity assay, RRB was higher than BRB, while BRB showed higher ${\alpha}$-glucosidase inhibitory activity than RRB. A strong and particular activity was observed in an anti-thrombosis activity assay of RRB. The extract of RRB demonstrated strong inhibitions against prothrombin and blood coagulation factors, with moderate thrombin inhibition. However, the extract of BRB did not exhibit any significant anti-thrombosis activity. Our results indicate that RRB has different, but useful biological activities, and loss or elimination of the biologically active substances in RRB occurs during the production of BRB. Therefore, to develop more functional foods from red beans, a study of suitable boiling, heating and drying processes is essential, and the efficient re-use of boiled waste water from the boiling process is necessary. These results could be applied to the further development of functional red bean beverages and sweat red bean pastes.

A Novel Glycine-Rich Region in Sox4 is a Target for the Proteolytic Cleavage in E. coli (전사활성 인자인 Sox4의 단백질 분해효소에 의한 표적 부위에 관한 연구)

  • 허은혜;최주연;장경희;김인경;임향숙
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.153-161
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    • 2002
  • Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

Inhibitory Effort of the N-terminal GST on the Tautomerase Activity of Macrophage Migration Inhibitory Factor (GST 융합 시스템에서 나타나는 macrophage migration inhibitory factor의 tautomerase 활성 저해에 관한 연구)

  • Kim Sang-Soo;Kim Kyung-Hee;Park Hyo-Jin;Hur Eun-hye;Rhim Hyangshuk
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.961-967
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    • 2005
  • Macrophage migration inhibitory fartor (MIF), known as a cytokine, is a multifunctional protein that is ubiquitously expressed in a variety of cells and tissues; however, enzymatic function of MIF still remains elusive in cells. In this study, we assessed details of the tautomerase activity of MIF. We established rapid purification condition for MIF by using pGEX system and compared the L-dopachrome tautomerase activity of GST-MIF, tMIF, and MIF. The results show that GST (glutathione S-transferase)-epitope tag or N-terminal amino acids flanking the essential $P^{2}$ almost completely abrogated L-dopachrome tautomerase activity of MIF. Subsequently, to determine whether the N-terminal tags have effects on oligomerization of MIF, protein cross-linking products were analyzed on $15\%$ SDS-PACE. The result demonstrates that N-terminal tags are dispensable for the formation of MIF's homooligomers. Thus, the results imply that exposure of If containing hydrophobic pocket in the active site is critical for L-dopachrome tautomerase activity of MIF. In addition, our study suggest that the MIF's tautomerase activity might be influenced by interacting with cellular partners.