• Title/Summary/Keyword: Biological activities

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Evaluation of Luminescent P450 Analysis for Directed Evolution of Human CYP4A11

  • Choi, Seunghye;Han, Songhee;Lee, Hwayoun;Chun, Young-Jin;Kim, Donghak
    • Biomolecules & Therapeutics
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    • v.21 no.6
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    • pp.487-492
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    • 2013
  • Cytochrome P450 4A11 (CYP4A11) is a fatty acid hydroxylase enzyme expressed in human liver. It catalyzes not only the hydroxylation of saturated and unsaturated fatty acids, but the conversion of arachidonic acid to 20-hydroxyeicosatetraenoic acid (20-HETE), a regulator of blood pressure. In this study, we performed a directed evolution analysis of CYP4A11 using the luminogenic assay system. A random mutant library of CYP4A11, in which mutations were made throughout the entire coding region, was screened with luciferase activity to detect the demethylation of luciferin-4A (2-[6-methoxyquinolin-2-yl]-4,5-dihydrothiazole-4-carboxylic acid) of CYP4A11 mutants in Escherichia coli. Consecutive rounds of random mutagenesis and screening yielded three improved CYP4A11 mutants, CP2600 (A24T/T263A), CP2601 (T263A), and CP2616 (A24T/T263A/V430E) with ~3-fold increase in whole cells and >10-fold increase in purified proteins on the luminescence assay. However, the steady state kinetic analysis for lauric acid hydroxylation showed the significant reductions in enzymatic activities in all three mutants. A mutant, CP2600, showed a 51% decrease in catalytic efficiency ($k_{cat}/K_m$) for lauric acid hydroxylation mainly due to an increase in $K_m$. CP2601 and CP2616 showed much greater reductions (>75%) in the catalytic efficiency due to both a decrease in $k_{cat}$ and an increase in Km. These decreased catalytic activities of CP2601 and CP2616 can be partially attributed to the changes in substrate affinities. These results suggest that the enzymatic activities of CYP4A11 mutants selected from directed evolution using a luminogenic P450 substrate may not demonstrate a direct correlation with the hydroxylation activities of lauric acid.

Comparison of Bioactive Constituents and Biological Activities of Aronia, Blackcurrant, and Maquiberry (아로니아, 블랙커런트, 마키베리의 기능성 성분 및 생리활성 비교)

  • Chung, Hai-Jung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.8
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    • pp.1122-1129
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    • 2016
  • In this study, bioactive constituents and biological activities of aronia, blackcurrant, and maquiberry were investigated. The 60% ethanol extracts were prepared and evaluated for total polyphenol, total flavonoid, and total anthocyanin contents. Biological activities, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing antioxidant power (FRAP), reducing power, tyrosinase inhibitory activity, and nitrite scavenging activity were also examined. Maquiberry showed the highest bioactive constituents as shown based on total polyphenol (73.66 mg GAE/g), total flavonoid (28.08 mg/g), and total anthocyanin (12.82 mg/g) contents compared to aronia and blackcurrant. Maquiberry also exhibited the highest DPPH and ABTS radical scavenging activity, FRAP, and reducing power at a concentration of 5 mg/mL, which were superior to those of ascorbic acid (a positive control). Therefore, maquiberry could be used as a potential source of antioxidants and functional food substances.

Discovery, semisynthesis, biological activities, and metabolism of ocotillol-type saponins

  • Liu, Juan;Xu, Yangrong;Yang, Jingjing;Wang, Wenzhi;Zhang, Jianqiang;Zhang, Renmei;Meng, Qingguo
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.373-378
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    • 2017
  • Ocotillol-type saponins are one kind of tetracyclic triterpenoids, sharing a tetrahydrofuran ring. Natural ocotillol-type saponins have been discovered in Panax quinquefolius L., Panax japonicus, Hana mina, and Vietnamese ginseng. In recent years, the semisynthesis of 20(S/R)-ocotillol-type saponins has been reported. The biological activities of ocotillol-type saponins include neuroprotective effect, antimyocardial ischemia, antiinflammatory, antibacterial, and antitumor activities. Owing to their chemical structure, pharmacological actions, and the stereoselective activity on antimyocardial ischemia, ocotillol-type saponins are subjected to extensive consideration. In this review, we sum up the discovery, semisynthesis, biological activities, and metabolism of ocotillol-type saponins.

Effects of Mixed Planting of Green Manure Crops Supplemented with Humic Substance on the Biological Soil Health Indicators of Reclaimed Soils (녹비식물 혼합재배에 휴믹물질 투입이 정화처리토양의 생물학적 토양 건강성 지표에 미치는 영향)

  • Bae, Bumhan;Park, Hyesun;Kang, Sua
    • Journal of Soil and Groundwater Environment
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    • v.26 no.5
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    • pp.49-59
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    • 2021
  • The effects of green manure crops, hairy vetch and sesban, supplemented with HS (humic substance) on biological soil health indicators was studied in a pot containing two kinds of reclaimed soil previously contaminated with petroleum hydrocarbons; a soil remediated by land-farming (DDC) and another soil by low-temperature thermal desorption (YJ). Treatments include no plant (C), plants only (H), and plants+2% HS (PH), which were evaluated in a pot containing respective soil. Biological indicators include microbial community analysis as well as soil enzyme activities of dehydrogenase, 𝛽-glucosidase, N-acetyl-𝛽-D-glucosaminidase (NAG), acid/alkaline phosphatase, arylsulfatase, and urease. Results showed an increase of enzyme activities in pot soils with plants and even greater in soils with plants+HS. The enzyme activities of DDC soil with plants (DDC_P) and with plants+HS (DDC_PH) increased 1.6 and 3.9 times on average, respectively than those in the control. The enzyme activities YJ soil with plants (YJ_P) and with plant+HS (YJ_PH) increased 1.8 and 3.8 times on average, respectively than those in the control. According to microbial community analysis, the relative abundance of nitrogen-fixing bacteria in DDC and YJ soil was increased from 1.5% to 7% and from 0 to 5%, respectively, after planting hairy vetch and sesban. This study showed that mixed planting of green manure crops with a supplement of humic substance is highly effective for the restoration of biological health indicators of reclaimed soils.

A Study on Biological Activities of Opuntia humifusa Cladode Extracts (손바닥 선인장 (Opuntia humifusa) 줄기 추출물의 생리활성)

  • Yoon, Min-Sun;Yoo, Jae-Soo;Lee, Keun-Kwang;Kim, Myung-Kon
    • Journal of Applied Biological Chemistry
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    • v.55 no.2
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    • pp.117-121
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    • 2012
  • Biological activities of the hot water and ethanol extracts from Opuntia humifusa cladodes were investigated. 1,1-diphenyl-2-picryl hadrazyl (DPPH) electron donating ability of hot water and ethanol extracts was 79.07 and 82.54%, respectively. Hot water extract generally showed better cytotoxic activity than ethanol extract against each cell line. HeLa and AGS cell lines treated with hot water extract had more than 50% cytotoxic activities. Based on the antimicrobial activities against four microbial strains, both extracts inhibited growth of Staphylococcus aureus KCCM 11593, whereas affected cell growth of three other microorganisms, Escherichia coli (KCCM 11234), Pseudomonas aeruginosa (ATCC 27853), and Salmonella typhimurium (ATCC 11862), in proportion to the concentration of extracts. The inflammatory activities against hot water extract (34.31%) showed higher than that of ethanol extract (25.59%). The effect of extracts on 3T3-L1 preadipocytes differentiation showed that differentiation of treated group with 80 and 100 ${\mu}g/mL$ of hot and ethanol extracts were increased more than treated group with isobutyl methyl xanthine (IBMX) + dexamethasone. These results indicate that the O. humifusa cladodes extracts can be used as a functional material due to their effective biological activities.

Various Biological Activities of Ramie (Boehmeria nivea)

  • Lee, Ah Young;Wang, Xiaoning;Lee, Dong Gu;Kim, Young-Mi;Jung, Yong-Su;Kim, Ho Bang;Kim, Hyun Young;Cho, Eun Ju;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • v.57 no.3
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    • pp.279-286
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    • 2014
  • The purpose of this study was to evaluate the biological activities of extracts of ramie (Boehmeria nivea (L.) Gaud.), hereafter referred to as Bn. Bn extracts from various collecting area were extracted with methanol. Two extracts from our study, Bn-13 and -82, showed significant antioxidant properties, likely due to their ability to scavenge free radicals. In addition, Bn extracts showed stronger anti-bacterial activity against Escherichia coli (Bn-40), Stapylococcus aureus (Bn-33), and Helicobacter pylori (Bn-05). In addition, this study was conducted to evaluate the anti-inflammatory effects of Bn extracts in lipopolyssacharide (LPS)- and interferon-${\gamma}$ (IFN-${\gamma}$)-stimulated RAW 264.7 macrophages cells. Bn-37 significantly inhibited the production LPS/IFN-${\gamma}$-induced nitric oxide. The most noteworthy anti-cancer effect was found in Bn-23. Bn-08 showed inhibition of aldose reductase. This study provides basic information for the development of functional foods.

Analysis of Biological Experiment on Socheongryong-tang (Xiaoqinglong-tang) (소청룡탕 효능에 관한 기초 실험 연구 문헌 분석)

  • Kim, Jung-Hoon;Shin, Hyeun-Kyoo
    • The Journal of Korean Medicine
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    • v.33 no.3
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    • pp.54-73
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    • 2012
  • Objectives: To investigate the scientific evidence of Korean Medicine (KM), papers on Socheongryong-tang (Xiaoqinglong-tang), which is frequently used in medical clinics or hospitals of Korean medicine, were collected and analyzed. Methods: Papers were classified by the registration of domestic or international journals, the year of publication, experimental models and the subjects of biological activities. The mechanisms of biological activity in accordance with therapeutic effects of Socheongryong-tang (Xiaoqinglong-tang) were noted. Results: Among 98 papers included, 21 were published in domestic journals whereas 35 were in Chinese journals and 43 in Japanese journals. Most reported biological activities were amelioration of asthma. Socheongryong-tang (Xiaoqinglong-tang) regulated interleukin and interferon and immunoglobulin, inhibited the production of nerve growth factor, endotheliln-1, nitric oxide, toll-like receptor-4, p-Akt and increased extracellular signal regulated kinase and cyclin D1, which led to decreased bronchi inflammation and bronchoconstriction, and inhibited the proliferation of airway smooth muscle cells, mucus secretion and airway hyperresponsiveness. In addition, Socheongryong-tang (Xiaoqinglong-tang) also restored tissues injured by asthma so that respiratory function recovered. Conclusions: Amelioration of asthma by Socheongryong-tang (Xiaoqinglong-tang) is supported by objective and scientific evidence.

Conformational change of organic cofactor PLP is essential for catalysis in PLP-dependent enzymes

  • Ngo, Ho-Phuong-Thuy;Nguyen, Diem Quynh;Park, Hyunjae;Park, Yoon Sik;Kwak, Kiwoong;Kim, Taejoon;Lee, Jang Ho;Cho, Kyoung Sang;Kang, Lin-Woo
    • BMB Reports
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    • v.55 no.9
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    • pp.439-446
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    • 2022
  • Pyridoxal 5'-phosphate (PLP)-dependent enzymes are ubiquitous, catalyzing various biochemical reactions of approximately 4% of all classified enzymatic activities. They transform amines and amino acids into important metabolites or signaling molecules and are important drug targets in many diseases. In the crystal structures of PLP-dependent enzymes, organic cofactor PLP showed diverse conformations depending on the catalytic step. The conformational change of PLP is essential in the catalytic mechanism. In the study, we review the sophisticated catalytic mechanism of PLP, especially in transaldimination reactions. Most drugs targeting PLP-dependent enzymes make a covalent bond to PLP with the transaldimination reaction. A detailed understanding of organic cofactor PLP will help develop a new drug against PLP-dependent enzymes.

Evaluation of Antioxidant and Antimicrobial Activities of Ethanol Extracts of Three Kinds of Strawberries

  • Seleshe, Semeneh;Lee, Jong Seok;Lee, Sarah;Lee, Hye Jin;Kim, Ga Ryun;Yeo, Joohong;Kim, Jong Yea;Kang, Suk Nam
    • Preventive Nutrition and Food Science
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    • v.22 no.3
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    • pp.203-210
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    • 2017
  • The antioxidant and antimicrobial activities of three kinds of strawberry ethanol extracts from Robus corchorifolius L. f. (RCL), Rubus parvifolius L. var. parvifolius (RPL), and Duchesnea chrysantha Miq. (DCM) were investigated. The RPL was highest (P<0.05) in phenolic, flavonoid, and anthocyanin contents. 2,2-Diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activities of RPL and DCM extracts were higher than that of RCL (P<0.05). Hydrogen peroxide scavenging activity of RPL was high compared to DCM and RCL (P<0.05). RCL exhibited a significant (P<0.05) potent antioxidant activity in nitric oxide radical inhibition. Inhibition diameter zone (nearest mm) of extracts against the test bacteria ranged from 11.5 in RCL to 12.5 in DCM against Staphylococcus aureus, from 10.5 in RCL to 13.5 in DCM against Streptococcus pneumoniae, from 8.5 in DCM to 10.5 in RCL against Escherichia coli, and the same inhibition of 10 mm in three of the extracts against Klebsiella pneumoniae. However, there was no inhibition against fungi Aspergillus niger and Candida albicans. Three of the extracts had the same minimum inhibitory concentration values of 12.50, 12.50, and $6.25{\mu}g/mL$ against S. aureus, K. pneumoniae, and S. pneumoniae, respectively. On the other hand, MIC values of 12.50, 12.50, and $6.50{\mu}g/mL$ were recorded for RPL, DCM, and RCL against E. coli, respectively. The result of present study revealed that extracts from three kinds of strawberries could be potential candidates as antioxidant and antimicrobial sources for functional food industries.