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Purification and Identification of Antioxidant Peptides from Enzymatic Hydrolysate of Spirulina platensis

  • Yu, Jie;Hu, Yuanliang;Xue, Mingxiong;Dun, Yaohao;Li, Shenao;Peng, Nan;Liang, Yunxiang;Zhao, Shumao
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1216-1223
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    • 2016
  • The aim of this study was to isolate antioxidant peptides from an enzymatic hydrolysate of Spirulina platensis. A novel antioxidant peptide was obtained by ultrafiltration, gel filtration chromatography, and reverse-phase high-performance liquid chromatography, with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay used to measure the antioxidant activity, and the sequence was determined to be Pro-Asn-Asn (343.15 Da) by electrospray ionization tandem mass spectrometry. This peptide was synthesized to confirm its antioxidant properties, and it exhibited 81.44 ± 0.43% DPPH scavenging activity at 100 μg/ml, which was similar to that of glutathione (82.63 ± 0.56%). Furthermore, the superoxide anion and hydroxyl free-radical scavenging activities and the SOD activity of the peptide were 47.84 ± 0.49%, 54.01 ± 0.82%, and 12.55 ± 0.75%, respectively, at 10 mg/ml. These results indicate that S. platensis is a good source of antioxidant peptides, and that its hydrolysate may have important applications in the pharmaceutical and food industries.

Soluble Expression and Purification of the Catalytic Domain of Human Vascular Endothelial Growth Factor Receptor 2 in Escherichia coli

  • Wei, Jia;Cao, Xiaodan;Zhou, Shengmin;Chen, Chao;Yu, Haijun;Zhou, Yao;Wang, Ping
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1227-1233
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    • 2015
  • Vascular endothelial growth factor (VEGF) plays a key role in angiogenesis through binding to its specific receptors, which mainly occurs to VEGF receptor 2 (VEGFR-2), a kinase insert domain-containing receptor. Therefore, the disruption of VEGFR-2 signaling provides a promising therapeutic approach for the treatment of cancer by inhibiting abnormal or tumorinduced angiogenesis. To explore this potential, we expressed the catalytic domain of VEGFR-2 (VEGFR-2-CD) as a soluble active kinase in Escherichia coli. The recombinant protein was purified and the VEGFR-2-CD activity was investigated. The obtained VEGFR-2-CD showed autophosphorylation activity and phosphate transfer activity comparable to the commercial enzyme. Furthermore, the IC50 value of known VEGFR-2 inhibitor was determined using the purified VEGFR-2-CD. These results indicated a possibility for functional and economical VEGFR-2-CD expression in E. coli to use for inhibitor screening.

Molecular Characterization of Cold-Inducible ${\beta}$-Galactosidase from Arthrobacter sp. ON14 Isolated from Antarctica

  • Xu, Ke;Tang, Xixiang;Gai, Yingbao;Mehmood, Muhammad Aamer;Xiao, Xiang;Wang, Fengping
    • Journal of Microbiology and Biotechnology
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    • v.21 no.3
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    • pp.236-242
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    • 2011
  • A psychrotrophic bacterium, Arthrobacter sp. ON14, isolated from Antarctica, was shown to exhibit a high ${\beta}$-galactosidase activity at a low temperature. A genomic library of ON14 was constructed and screened for ${\beta}$-galactosidase genes on functional plates containing 5-bromo-4-chloro-3-indolyl-${\beta}$-D-galactopyranoside (X-gal) as the substrate. Two different ${\beta}$-galactosidase genes, named as galA, galB, were found in ON14. Computational analyses of the genes revealed that the encoded protein GalA belongs to family 2 of glycosyl hydrolysases and is a cold-active protein, whereas GalB belongs to family 42 of glycosyl hydrolysases and is a mesophilic protein. Reverse transcription analyses revealed that the expression of galA is highly induced at a low temperature ($4^{\circ}C$ ) and repressed at a high temperature ($28^{\circ}C$ ) when lactose is used as the sole carbon source. Conversely, the expression of galB is inhibited at a low temperature and induced at a high temperature. The purified GalA showed its peak activity at $15^{\circ}C$ and pH 8. The mineral ions $Na^+$, $K^+$, $Mg^{2+}$, and $Mn^{2+}$ were identified as enzyme activators, whereas $Ca^{2+}$ had no influence on the enzyme activity. An enzyme stability assay revealed that the activity of GalA is significantly decreased when it is incubated at $45^{\circ}C$ for 2 h, and all its activity is lost when it is incubated at $50^{\circ}C$.

Purification and Characterization of an Antilisterial Bacteriocin Produced by Leuconostoc sp. W65

  • Oh, Se-Jong;Kim, Myung-Hee;Churey, John-J.;Worobo, Randy-W.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.680-686
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    • 2003
  • This study was carried out to characterize the antilisterial substances produced by Leuconostoc sp. W65 and to evaluate the effects of pH, temperature, and time on inhibitory activity using response surface methodology. Leucocin W65, an antilisterial substance produced by Leuconostoc sp. W65, markedly inhibited the growth of Listeria monocytogenes, L. innocua, and L. ivanovii, whereas other pathogens including Gram-negative bacteria were not susceptible. The pH was the most effective factor with regard to bacteriocin activity, while temperature and time of heat treatment had no significant effect. Fifty percent of inhibitory activity remained after 22.8 min at pH 4.2 and $121^{\circ}C$. Leucocin W65 was purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and tricine-SDS-PAGE. Compositional analysis originally estimated the peptide to be 56 amino acids in length without asparagine, glutamine, and tryptophane. The sequence of partial N-terminal amino acid residues of purified bacteriocin was identified as follows: $NH_{2}-XGXAGVXPXGGQQPXVPLXYP$.

Effect of metal ions on the secondary structure and activity of calf intestine phosphatase

  • Chen, Fengjuan;Liu, Guoqi;Xu, Zhihong;Zeng, Zhengzhi
    • BMB Reports
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    • v.41 no.4
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    • pp.305-309
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    • 2008
  • Cobalt is an essential microelements in many biological processes involving enzymatic activity. We found that $Zn^{2+}$ and $Mg^{2+}$, which are in the active site of native calf intestine alkaline phosphatase (CIP), can be replaced by $Co^{2+}$ directly in solution. The effect of $Co^{2+}$ concentration on the substitution reaction was examined at ratios of [$Co^{2+}$]/[CIP] from 0:1 to 8:1. The quantity of $Zn^{2+}$ in CIP decreased progressively as the ratio was increased, but the amount of $Mg^{2+}$ changed in irrregular fashion. A series of active site models of the reaction mechanism of CIP are proposed. Low pH was found to promote the replacement of $Mg^{2+}$ by $Co^{2+}$. To understand how the substitution affects the enzyme, we also solved the secondary structure of CIP after reaction with $Co^{2+}$ in different conditions.

Influence of Glycyrrhizic Acid, Menthol and Their Supramolecular Compounds on the Functional Activity of Rat Mitochondria in in-vitro Experiments

  • Ettibaeva, L.A.;Abdurahmonova, U.K.;Matchanov, A.D.;Allanazarova, D.M.;Halmuratova, Z.T.
    • Journal of the Korean Chemical Society
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    • v.65 no.5
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    • pp.313-319
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    • 2021
  • Menthol (M) is a cyclic monoterpenode and is a major component of essential oils. Menthol, along with menthol, isomenton, etc., gives taste and odor of the mint plant, and when it comes to menthol in general, L- or (-) -menthol is usually used. Included in pharmaceuticals, cosmetics and pesticides. It has antimicrobial, antibacterial, antioxidant properties. It is also known that the licorice plant (Glycyrrhiza Glabra L.) differs from other types of plants by its medicinal properties. For many years it has been used in folk medicine. Extraction of licorice root revealed up to 25% glycyrrhizinic acid (GA). Its aglycone - glycyrrhizic acid is notable for its structural similarity to the adrenal cortex hormones. Currently, GA and glycyrrhizic acid are widely used in medicine as a remedy for colds, allergies, viral diseases, tumors. The biological activity of menthol and GA-based supramolecular compounds has been poorly studied, and their effect on the functional parameters of rat liver mitochondria has been studied little. For this purpose, in our experiments, the effect of menthol (M), glycyrrhizinic acid (GA) and their supramolecular complexes obtained in different proportions on in vitro and in vivo studies on rat liver mitochondria was studied.

A Study on the Influence of Knowledge Management System Quality on the Work Performance of Public Institutions: Focusing on the Interaction between Collective Intelligence and Innovative Activities (공공기관의 지식관리시스템 품질이 업무성과에 미치는 영향에 관한 연구: 집단지성과 혁신활동의 상호작용을 중심으로)

  • Yu, Dalyoung;Park, Sungbum
    • Journal of Information Technology Services
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    • v.21 no.1
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    • pp.103-125
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    • 2022
  • This study empirically examines the correlation between the data, system, service quality, collective intelligence, innovation activity, and work performance of the KMS for employees of state-run enterprises. This study also confirmed that collective intelligence and innovation activity have a mediating effect on the relationship between KMS system quality and work performance. For this study, employees of state-run enterprises completed a 5-point Likert scale survey, and the data obtained were analyzed with descriptive statistics. The reliability and validity of variables were also verified through a measurement model analysis. Structural equation modeling of IBM SPSS 26 and AMOS 26 were used to prove the hypothesis. Through this study, For the improvement of sustainable work performance of state-run enterprises, which is a difficult issue to solve, not only should the quality of the KMS utilized in the enterprises be improved and managed, but also the design for various incentive schemes tailored to the enterprises' job characteristics, such as rewards, HR bonus, work incentives, job allowance, and encouragement from the head of the enterprise, should be reviewed and continuously developed in reflection of the active collective intelligence of the employees and the effect of innovation activities. Then, the work performance of public institutions will increase.

Regulation of Nrf2 Transactivation Domain Activity by p160 RAC3/SRC3 and Other Nuclear Co-Regulators

  • Lin, Wen;Shen, Guoxiang;Yuan, Xiaoling;Jain, Mohit R.;Yu, Siwang;Zhang, Aihua;Chen, J. Don;Kong, Ah-Ng Tony
    • BMB Reports
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    • v.39 no.3
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    • pp.304-310
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    • 2006
  • Transcription factor NF-E2-related factor 2 (Nrf2) regulates the induction of Phase II detoxifying enzymes and antioxidant enzymes in response to many cancer chemopreventive compounds. In this study, we investigated the role of receptor associated coactivator (RAC3) or steroid receptor coactivator-3 (SRC3) and other nuclear co-regulators including CBP/p300 (CREB-binding protein), CARM1 (Coactivator-associated arginine methyltransferase), PRMT1 (Protein arginine methyl-transferase 1), and p/CAF (p300/CBP-associated factor) in the transcriptional activation of a chimeric Gal4-Nrf2-Luciferase system containing the transactivation domain (TAD) of Nrf2 in HepG2 cells. The results indicated that RAC3 up-regulated the transactivation activity of Gal4-Nrf2-(1-370) in a dose-dependent manner. The enhancement of transactivation domain activity of Gal4-Nrf2-(1-370) by RAC3 was dampened in the presence of dominant negative mutants of RAC3. Next we studied the effects of other nuclear co-regulators including CBP/p300, CARM1, PRMT1 and p/CAF, and the results showed that they had different level of positive effects on this transactivation domain activity of Gal4-Nrf2-(1-370). But importantly, synergistic effects of these co-regulators in the presence of RAC3/SRC3 on the transactivation activity of Gal4-Nrf2-(1-370) were observed. In summary, our present study showed for the first time that the 160 RAC3/SRC3 is involved in the functional transactivation of TAD of Nrf2 and that the other nuclear co-regulators such as CBP/p300, CARM1, PRMT1 and p/CAF can also transcriptionally activate this TAD of Nrf2 and that they could further enhance the transactivation activity mediated by RAC3/SRC3.

Effects of Cadmium and Cadmium Binding Protein on the Respiration of Mitochondria of Rat Kidney (흰쥐 신장의 미토콘드리아 호흡에 미치는 Cadmium과 Cadmium Binding Protein의 영향)

  • Cho, Hae-Kyoung;Boo, Moon-Jong;Choe, Rim-Soon
    • Applied Microscopy
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    • v.19 no.2
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    • pp.59-73
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    • 1989
  • To investigate the effect of cadmium and cadmium binding protein on the electron transport system and conformational changes of rat kidney mitochondria, various cadmium concentration were treated in vitro and respiration rate, NADH-CoQ reductase activity were measured. Ultrastructural changes at state IV respiration were also observed. CdBP was isolated from the rat liver by Sephadex G-75 column fractionation and treated in vitro with cadmium. Also mitochondrial state IV respiration rate was measured. When cadmium was treated in vitro, state IV respiration and enzyme activity were decreased and ultrastructural transformation of mitochondria from a condensed to an orthodox conformation was inhibited under state IV respiration. In case cadmium and CdBP were treated together, oxygen consumption was more increased than cadmium only. Conformational changes of mitochondria from a condensed to orthodox conformation were also observed. This indicates that CdBP have a protective effect against cadmium toxicity.

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On the Application of Public Search Measures to Detect and Obtain Information

  • Kozytska, Olena;Tsilmak, Olena;Protsenko, Olena;Yankovyi, Mykola;Lysenko, Аndrii;Shulzhenko, Assol
    • International Journal of Computer Science & Network Security
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    • v.21 no.9
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    • pp.109-112
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    • 2021
  • The article considers the state of legislation that regulates the use of public methods of obtaining information by authorized state bodies. The correlation of public investigative (search) actions with operative-search measures as concepts denoting the application of public methods of obtaining information has been studied. In addition, it argues the need for more detailed delineation and legislative regulation of public investigative (search) actions and operational and investigative measures at the legislative and departmental levels. The purpose of the article is to analyze certain provisions of the Law of Ukraine "On operational and investigative activities" to identify inconsistencies in the content of the text of the law in order to correct and prevent ambiguity in the theory and practice of law enforcement.