Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Cloning and Characterization of Flavone synthase I from Populus deltoids

포플러로부터 flavone synthase I 유전자의 클로닝 및 생화학적 특성

  • Kim, Bong-Gyu (Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University) ;
  • Ahn, Joong-Hoon (Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University)
  • 김봉규 (건국대학교 생명공학과, 건국대학교 생명분자정보학 센터) ;
  • 안중훈 (건국대학교 생명공학과, 건국대학교 생명분자정보학 센터)
  • Published : 2009.03.31
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Abstract

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, $Fe^{2+}$, ascorbate and catalase. Thus, it is concluded that PFNS N-1 encodes a flavone synthase I.

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 이루어지지 않았다. 포플러에서 flavone synthase I 유전자를 RT-PCR방법을 이용하여 클로닝하였다. 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, $FeSO_4$, 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.

Keywords

References

  1. Britsch L and Grisebach H (1986) Purification and characterization of (2S)-flavanone 3-hydroxylase from Petunia hybrid. Eur J Biochem 156, 569-577 https://doi.org/10.1111/j.1432-1033.1986.tb09616.x
  2. Cervera M-T, Storme V, Ivens B, Gusmao J, Liu BH, Hostyn V, Van Slycken J, Van Montagu M, and Boerjan W (2001) Dense genetic linkage maps of three Populus species (Populus deltoides, P. nigra and P. trichocarpa) based on AFLP and microsatellite markers. Genetics 158, 787-809
  3. Christensen AB, Gregersen PL, Olsen CE, and Collinge DB (1998) A flavonoid 7-O-methyltransferase is expressed in barley leaves in response to pathogen attack. Plant Mol BioI 36, 219-227 https://doi.org/10.1023/A:1005985609313
  4. Dixon RA, Achnine L, Kota P, Liu C-J, Srinivasa Reddy MS, and Wang L (2002) The phenylpropanoid pathway and plant defense - a genomics perspective. Mol Plant Pathol 3, 371-390 https://doi.org/10.1046/j.1364-3703.2002.00131.x
  5. Forkmann G and Heller W (1999) Biosynthesis of flavonoids. In: Barton D, Nakanishi K, and Meth-Cohn O (eds.), Comprehensive Natural Products Chemistry. Elsevier Science Ltd., Oxford, pp. 713-748
  6. Harborne JB and Williams C (2000) Advances in flavonoid research since 1992. Phytochemistry 55, 481-504 https://doi.org/10.1016/S0031-9422(00)00235-1
  7. Havsteen B (2003) Flavonoids, a class of natural products of high pharmacological potency. Biochem Pharmaco 32, 1141-1148 https://doi.org/10.1016/0006-2952(83)90262-9
  8. Heller W and Forkmann G (1993) Biosynthesis of flavonoids. In: Harborne JB (ed.), The Flavonoids: Advances in Research since 1986. Chapman & Hall, London, pp. 499-535
  9. Herve P, Jauneau A, Paques M, Marien IN, Boudet AM, and Teulieres C (2001) A procedure for shoot organogenesis in vitro from leaves and nodes of an elite Eucalyptus gunnii clone: Comparative histology. Plant Sci 161, 645-653 https://doi.org/10.1016/S0168-9452(01)00451-4
  10. Katsuyama Y, Funa N, Muyahisa I, and Horinouchi S (2007) Synthesis of unnatural flavonoids and stilbenes by exploiting the plant biosynthetic pathway in Escherichia coil. Chem BioI 14, 613-621 https://doi.org/10.1016/j.chembiol.2007.05.004
  11. Kim BG, Kim JH, Kim J, Lee C, and Ahn J-H (2008) Accumulation of flavonols in response to ultraviolet-B irradiation in soybean is related to induction of flavanone 3-$\beta$-hydroxylase and flavonol synthase. Mol Cells 25, 247-252
  12. Kim BG, Lee Y, Hur H-G, Lim Y, and Ahn J-H (2006) Flavonoid 3'-O-methyltransferase from rice: cDNA cloning, characterization and functional expression. Phytochemistry 67, 387-394 https://doi.org/10.1016/j.phytochem.2005.11.022
  13. Lee YJ, Kim JH, Kim BG, Lim Y, and Ahn J-H (2008) Characterization of flavone synthase I from rice. BMB Rep 41, 68-71
  14. Leonard E, Yan Y, Fowler ZL, Li Z, Lim C-G, Lim K-H, and Koffas MAG (2008) Strain improvement of recombinant Escherichia coli for efficient production of plant flavonoids. Mol Pharm 5, 257-265 https://doi.org/10.1021/mp7001472
  15. Li L, Cheng XF, Leshkevich J, Umezawa T, Harding SA, and Chiang VL (2001) The last step of syringyl monolignol biosynthesis in angiosperms is regulated by a novel gene encoding sinapyl alcohol dehydrogenase. Plant Cell 13, 1567-1585 https://doi.org/10.1105/tpc.13.7.1567
  16. Martens S, Forkmann G, Matern U, and Lukaein R (2001) Cloning of parsley flavone synthase I. Phytochemistry 58, 43-46 https://doi.org/10.1016/S0031-9422(01)00191-1
  17. Martens S and Mithofer A (2005) Flavones and flavone synthases. Phytochemistry 66, 2399-2407 https://doi.org/10.1016/j.phytochem.2005.07.013
  18. Miyahisa I, Kaneko M, Funa N, Kawasaki H, Kojima H, Ohnishi Y, and Horinouchi S (2006) Efficient production of (2S)flavones by Escherichia coli contammg an artificial biosynthetic gene cluster. Appl Microbiol Biotechnol 68, 498-504 https://doi.org/10.1007/s00253-005-1916-3
  19. Schijlen EG, de Vos RCH, van Tunen AJ, Bovy AG (2004) Modification of flavonoid biosynthesis in crop plants. Phytochemistry 65, 2631-2648 https://doi.org/10.1016/j.phytochem.2004.07.028
  20. Tahara S (2007) A journey of twenty-five years through the ecological biochemistry of flavonoids. Biosci Biotechnol Biochem 71, 1387-1404 https://doi.org/10.1271/bbb.70028
  21. Tsai C-J, Harding S, Tschaplinski T, Lindroth R, and Yuan Y (2006) Genome-wide analysis of the structural genes regulating defense phynylpropanoid metabolism in Populus. New Phytologist 172, 47-62 https://doi.org/10.1111/j.1469-8137.2006.01798.x
  22. Turnbull JJ, Nakajima J, Welford RWD, Yamazaki M, Saito K, and Schorield CJ (2004) Mechanistic studies on three 2-oxoglutaratedependent oxygenases of flavonoid biosynthesis. J BioI Chem 279, 1206-1216 https://doi.org/10.1074/jbc.M309228200
  23. Tuskan GA, DiFazio S, Jansson S, Bohlmann J, Grigoriev I, Hellsten U, Putnam N, Ralph S, Rombauts S, Salamov A et al. (2006) The genome of black cottonwood, Populus trichocarpa (Torr. & Gray). Science 313, 1596-1604 https://doi.org/10.1126/science.1128691
  24. Winkel-Shirley B (2001) Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. Plant Physiol 126, 485-493 https://doi.org/10.1104/pp.126.2.485
  25. Yin TM, Difazio SP, Gunter LE, Riemenschneider D, and Tuskan GA (2004) Large-scale heterospecific segregation distortion in Populus revealed by a dense genetic map. Theor Appl Genet 109, 451-463

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