포플러로부터 flavone synthase I 유전자의 클로닝 및 생화학적 특성

김봉규,안중훈 한국응용생명화학회 2009 Journal of Applied Biological Chemistry (J. Appl. Vol.52 No.1

포플러는 naringenin, kaempferol, myricetin, apigenin, luteolin, rhamnetin, quercetin과 같은 플라보노이드를 가지고 있다. 이러한 플라보노이드들은 여러 가지 효소에 의해 naringenin으로부터 합성된다. 그러나, 포플러에서는 플라보노이드 합성에 관여하는 효소들의 연구가 거의 이루어 지지 않았다. 포플러 에서 flavone synthase I 유전자를 RT-PCR방법을 이용하여 클로닝 하였다. PFNS I-1 유전자의 기질 특성을 알아보기 위하여 대장균에 과발현하여 glutathione S-transferase 친화 크로마토그래피를 이용하여 정제하였다. 정제한 PFNS I-1 재조합 단백질은 flavanone인 2S-naringenin기질을 이용하여 flavone인 apigenin을 생성함을 알 수 있었다. 또한 cofactor인 oxoglutarate, FeSO4, ascorbate, catalase가 PFNS I-1의 반응성에 중대한 영향을 미치는 것을 알 수 있었다. 이상의 결과를 종합해 볼 때 PFNS I-1 유전자는 flavone synthase I을 암호화하는 유전자임을 확인하였다. Poplar contains various flavonoids including naringenin, kaempferol, myricetin, apigenin, luteolin,rhamnetin, and quercetin. These flavonoids are synthesized from naringenin with various enzymes. However, none of genes from poplar involved in flavonoid biosynthesis have been biochemically characterized. We cloned PFNS I-1 from Populus deltoids by RT-PCR method. The open reading frame of PFNS I-1 consisted of 1,017-bp and it showed high similarity with other FNS genes. The purified recombinant PFNS I-1, expressed in Escherichia coli, catalyzed the reaction from flavanone (naringenin) to flavone (apigenin). The reaction of PFNS I-1 was enhanced by cofactors such as oxoglutarate, Fe2+, ascorbate and catalase. Thus, it is concluded that PFNS I-1 encodes a flavone synthase I.

Antioxidant Properties of Flavone-6(4')-Carboxaldehyde Oxime Ether Derivatives

Gulgun, Ayhan-Kilcigil,Coban, Tulay,Tuncbilek, Meral,Benay, Can-Eke,Oya, Bozda-Dundar,Ertan, Rahmiye,Iscan, Mumtaz The Pharmaceutical Society of Korea 2004 Archives of Pharmacal Research Vol.27 No.6

The in vitro antioxidant properties of some flavone-6(4)-carboxaldehyde oxime ether deriva-tives (Ia-f, lIa-f) were determined by their effects on the rat liver microsomal NADPH-dependent lipid peroxidation (LP) levels by measuring the formation of 2-thiobarbituric acid reactive substances. The free radical scavenging properties of the compounds were also examined in vitro by determining their capacity to scavenge superoxide anions and interact with the stable free radical 2, 2-diphenyl-1-picrylhydrazyl (DPPH). The most active compounds, lib (Flavone-4'-carboxaldehyde-O-ethyl oxime) and Id (Flavone-6-carboxaldehyde-O-[2-(1-pyrolidino) ethyl] oxime), caused 98 and 79% inhibition of superoxide anion production and DPPH stable free radical at $10^{-3}{\;}M$, respectively.

Flavone의 합성 중간체인 2-Aroyloxyacetophenone류의 새로운 합성법

송규용,안병준 충남대학교 약학대학 의약품개발연구소 1994 藥學論文集 Vol.10 No.-

We have synthesized some 2-(2-benzyloxybenzoyloxy)-and 2-(2,6-dibenzyloxybenzoyloxy)-acetophenones as intermediates for flavone synthesis. The reaction of polyoxygenated 2-hydroxyacetophenones with 2-benzyloxybenzoic acid or 2,6-dibenzyloxybenzoic acid in the presence of dicyclohexylcarbodiimide and p-dimethylaminopyridine resulted in a good yield(70~89%) of 2-(2-benzyloxybenzoyloxy)-acetophenones or 2-(2,6-dibenzyloxybenzoyloxy)-acetophenones under milder conditions and in shorter time than the previous methods. This new methods using benzoic acids instead of benzoyl chlorides saves one reaction step of acid chlorination in comparision of the previous methods.

Pentahydroxy flavone에 依한 Vanadium(V)의 抽出吸光度定量

金永洙,趙誠烋,鄭鍾南 조선대학교 생산기술연구소 1979 生産技術硏究 Vol.1979 No.-

A brownish Vanadium(V)-pentahydroxy flavone chelate showed an absorption maximum at 520 mμ with a constant absorbance at PH 3―4.5, and it was applied to the determination of small amounts of Vanadiumt(V). The chelate was extracted with isoamyl alcohol. The chelate was as follows: After the sample solution containg Vanadium(V) was heated at 70―80℃ for 30 min. in an acetate buffer solution(PH 3-4.5), 5㎖ of iso-amylalcohol and 1.O㎖ of 0.04M-pentahydroxy flavone solution) in dimethylformamide) were added. The complex was extracted into organic phase by shaking for about 10 min., and absorbance at 520mμ was measured after one hour. In the concentration range of 5-100 of Vanadium (V) in 5㎖ of organic phase the calibration curve obeyed the Beer's Law. Co(II), Ni(II), Cu(II), Zn(II), Mn(II), Al(II), Mo(VI), etc. scarcely interfered even when their amounts were 200-500μg. Fe(III) and W(VI) interfered with the determination.

Bioconversion of Flavones During Fermentation in Milk Containing Scutellaria baicalensis Extract by Lactobacillus brevis

( Chen Xu ),( Geun Eog Ji ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.10

Scutellaria baicalensis (SB), a traditional herb with high pharmacological value, contains more than 10% flavone by weight. To improve the biological activity of flavones in SB, we aimed to enhance the bioconversion of baicalin (BG) to baicalein (B) and wogonoside (WG) to wogonin (W) in SB during fermentation using beta-glucuronidase produced from Lactobacillus brevis RO1. After activation, L. brevis RO1 was cultured in milk containing SB root extract with various carbon or nitrogen sources at 37oC for 72 h. During fermentation, the growth patterns of L. brevis RO1 and changes in the flavone content were assessed using thin-layer chromatography and high-performance liquid chromatography. After 72 h of fermentation, the concentrations of B and W in the control group increased by only 0.15 and 0.12 mM, respectively, whereas they increased by 0.57 and 0.24 mM in the fish peptone group. The production of B and W was enhanced by the addition of 0.4% fish peptone, which not only improved the growth of L. brevis RO1 (p < 0.001) but also enhanced the bioconversion of flavones. In conclusion, the bioconversion of flavones in SB may provide a potential application for the enhancement of the functional components in SB.

무궁화 품종내의 flavonoid 성분분포에 관한연구

유기역 한국자원식물학회 1996 한국자원식물학회지 Vol.9 No.3

무궁화 품종 6계통, 총 68종류를 선발하여 flavonoid 성분을 비교하였다. 2차원으로 크로마토그래피를 전개 후 13개의 flavonoid가 관찰되었고 spot 5번의 flavonoid 함량이 전체의 50%를 차지하였으며 나머지는 미량으로 관찰되었다. 6개의 group별 spot출현 양상은 각각 차이를 보였으나 group 내에서는 거의 동일한 것으로 나타났다. 13개의 spot 중 1, 4, 6은 flavonol의 특성을 나타냈으며 나머지 10개의 spot들이 flavone의 특성을 나타내었다. 품종별 flavone함량 및 분포는 백색계통이나 자주 및 청색 계통에는 많은 양이 존재 하였고 pink와 적색 group에서는 매우 적은 양이 분포하였다. Purple spot들의 flavonoid aglycone은 4’, 5-OH의 aglycone 특성을 보였으며 이들 중 spot 5는 saponarin, 7은 vitexin, 9는 xylovitexin, 11은 rhamnosylvitexin으로 동정되었으며 13은 apigenin-7-O-diglycoside로 추정되었다. Saponarin은 백색무궁화에서 전체 flavone량의 50% 이상을 점유하였다. Floral flavonoids of Hibiscus syriacus L. six complex with 68 formac all in all were examined. Thirteen flavonoids appeared on the two dimensional chromatogtams. Spot 5, however, occupied more than 50% in total flavonoid contents, and other spots were invariably minor pigments in all samples examined. Ten spots among 13 spots showed the characteristics of flavones, having color of purple to dark purple under UV light and yellow under ammonia gas, while spots reagents suggests that 10 purple spots are 4', 5-OH aglycone type. Four spots out of 10 purple spots were possible to be identified: spot 5, saponarin, spot 7, vitexin, spot 9, xylovitexin, and spot 11, rhamnosylvitexin, respectively. It was suggested that spot 13 might be apigenin-7-O-diglycoside.

Structural Characterization of a Flavonoid Compound Scavenging Superoxide Anion Radical Isolated from Capsella bursa-pastoris

Kweon, Mee-Hyang,Kwak, Jae-Hyock,Ra, Kyung-Soo,Sung, Ha-Chin,Yang, Han-Chul Korean Society for Biochemistry and Molecular Biol 1996 Journal of biochemistry and molecular biology Vol.29 No.5

A superoxide anion radical scavenger isolated from Capsella bursa-pastoris was characterized by infrared (IR) spectroscopy, sugar analysis, ultraviolet (UV) spectroscopy, $^{1}H$ and $^{13}C$ nuclear magnetic resonance (NMR) spectroscopies, and fast atom bombardment (FAB) mass analysis. The compound was assumed to be a flavonoid-O-glycoside from IR spectrum and UV absorption maxima. When the sugar composition of the compound was examined by thin layer chromatography (TLC) and gas chromatography (GC) of the acid hydrolysate, only glucose was detected. According to the results of UV spectrotroscopy by using shift reagents, the compound was supposed to be luteolin (5,7,3',4'-tetrahydroxy flavone) or chrysoeriol (5,7,4'-trihydroxy-3'-methoxy flavone) with glucose. Based on $^{1}H$- and $^{13}C-NMR$ spectroscopies, the compound was deduced as 7,4'-dihydroxy-5,3'-dimethoxy-${\alpha}$-6-c-glucosyl-${\beta}$-2"-o-glucosyl flavone. In FAB mass analysis the compound was finally characterized as 7,4'-dihydroxy-5,3'-dimethoxy-${\alpha}$-6-c-glucosyl-${\beta}$-2"-o-glucosyl flavone ($C_{29}H_{34}O_{16}$, M.W.=638).

Aqueous Solubility Enhancement of Some Flavones by Complexation with Cyclodextrins

김현명,정선호,김현원 대한화학회 2008 Bulletin of the Korean Chemical Society Vol.29 No.3

The inclusion complexes of cyclodextrins (CDs) with flavones in aqueous solution were investigated by phase solubility measurements. The effect of b -cyclodextrin (b -CD), heptakis (2,6-di-O-methyl) b -cyclodextrin (DM-b -CD) and 2-hydroxypropyl-b -cyclodextrin (HP-b -CD) on the aqueous solubility of three flavones, namely, chrysin, apigenin and luteolin was investigated, respectively. Solubility enhancements of all flavones obtained with three CDs followed the rank order: HP-b -CD > DM-b -CD > b -CD, and besides, CDs show higher stability constant on luteolin than that on others flavones. 1H-nuclear magnetic resonance (NMR) spectroscopy and molecular modeling was used to help establish the model of interaction of the CDs with luteolin. NMR spectroscopic analysis suggested that A-C ring, and part of the B ring of luteolin display favorable interaction with the CDs, which was also confirmed by docking studies based on the molecular simulation. The observed augmentation of solubility of luteolin by three CDs was explained by the difference of electrostatic interaction of each complex, especially hydrogen bonding.

Aqueous Solubility Enhancement of Some Flavones by Complexation with Cyclodextrins

Kim, Hyun-Myung,Kim, Hyun-Won,Jung, Seun-Ho Korean Chemical Society 2008 Bulletin of the Korean Chemical Society Vol.29 No.3

The inclusion complexes of cyclodextrins (CDs) with flavones in aqueous solution were investigated by phase solubility measurements. The effect of b -cyclodextrin (b -CD), heptakis (2,6-di-O-methyl) b -cyclodextrin (DM-b -CD) and 2-hydroxypropyl-b -cyclodextrin (HP-b -CD) on the aqueous solubility of three flavones, namely, chrysin, apigenin and luteolin was investigated, respectively. Solubility enhancements of all flavones obtained with three CDs followed the rank order: HP-b -CD > DM-b -CD > b -CD, and besides, CDs show higher stability constant on luteolin than that on others flavones. 1H-nuclear magnetic resonance (NMR) spectroscopy and molecular modeling was used to help establish the model of interaction of the CDs with luteolin. NMR spectroscopic analysis suggested that A-C ring, and part of the B ring of luteolin display favorable interaction with the CDs, which was also confirmed by docking studies based on the molecular simulation. The observed augmentation of solubility of luteolin by three CDs was explained by the difference of electrostatic interaction of each complex, especially hydrogen bonding.

Angelica keiskei의 고지혈증 개선효과 및 2종의 플라본 7-O-배당체의 분리

박종철,조영숙,박석규,박정로,전순실,옥광대,최종원,Park, Jong-Cheol,Cho, Young-Sook,Park, Seok-Kyu,Park, Jeong-Ro,Chun, Soon-Sil,Ok, Kwang-Dae,Choi, Jong-Won 한국생약학회 1995 생약학회지 Vol.26 No.4

Two flavone glycosides were isolated from the aerial part of Angelica keiskei Koidz and identified by means of spectral analysis as luteolin-$7-O-{\beta}-{_D}-glucoside$ and Iuteolin-7-O-rutinoside. Intraperitoneal administration of methanolic extract(100mg/kg) and $luteolin-7-O-{\beta}-{_D}-glucoside(5mg/kg)$ isolated from this plant produced a significant hypolipemic activity.