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Cytochrome P450S and Uses Thereof Joe Chappell University of Kentucky, [email protected]

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Cytochrome P450S and Uses Thereof Joe Chappell University of Kentucky, Chappell@Uky.Edu University of Kentucky UKnowledge Plant and Soil Sciences Faculty Patents Plant and Soil Sciences 9-11-2012 Cytochrome P450S and Uses Thereof Joe Chappell University of Kentucky, [email protected] Lyle F. Ralston Right click to open a feedback form in a new tab to let us know how this document benefits oy u. Follow this and additional works at: https://uknowledge.uky.edu/pss_patents Part of the Plant Sciences Commons Recommended Citation Chappell, Joe and Ralston, Lyle F., "Cytochrome P450S and Uses Thereof" (2012). Plant and Soil Sciences Faculty Patents. 9. https://uknowledge.uky.edu/pss_patents/9 This Patent is brought to you for free and open access by the Plant and Soil Sciences at UKnowledge. It has been accepted for inclusion in Plant and Soil Sciences Faculty Patents by an authorized administrator of UKnowledge. For more information, please contact [email protected]. US008263362B2 (12) United States Patent (10) Patent N0.: US 8,263,362 B2 Chappell et a]. (45) Date of Patent: *Sep. 11, 2012 (54) CYTOCHROME P450S AND USES THEREOF 2003/0166255 A1 9/2003 Chappell .................. .. 435/2523 2004/0078840 A1 4/2004 Chappell 6151. .. .. 800/278 2006/0218661 A1 9/2006 Chappell 6151. .. .. 800/278 (75) Inventors: Joseph Chappell, Lexington, KY (US); 2007/0231861 A1 10/2007 Millis et a1. 435/69.l Lyle F. Ralston, Kirkwood, MO (US) 2007/0238157 A1 10/2007 Millis et a1. .. 435/166 2007/0238159 A1 10/2007 Millis et a1. 435/25233 (73) Assignee: University of Kentucky Research 2007/0238160 A1 10/2007 Millis et a1. 435/25233 Foundation, Lexington, KY (US) 2007/0254354 A1 11/2007 Millis et a1. 435/25233 2008/0178354 A1 7/2008 Chappell 6151. 800/298 2008/0233622 A1 9/2008 Julien et a1. .. 435/148 (*) Notice: Subject to any disclaimer, the term of this 2010/0035329 A1 2/2010 Millis et a1. .. 435/2542 patent is extended or adjusted under 35 2010/0151519 A1 6/2010 Julien et a1. 435/69.l U.S.C. 154(b) by 32 days. 2010/0151555 A1 6/2010 Julien et a1. .. 435/193 2010/0216186 A1 8/2010 Chappell 6151. .. 435/69.l This patent is subject to a terminal dis 2011/0081703 A1 4/2011 Chappell 6151. ............ .. 435/193 claimer. FOREIGN PATENT DOCUMENTS (21) App1.No.: 12/182,000 JP 2000-511404 9/2000 wo 96/36697 11/1996 Filed: Jul. 29, 2008 wo 97/38571 10/1997 (22) wo 97/38703 10/1997 wo wo 97/37664 10/1997 (65) Prior Publication Data wo 00/17327 3/2000 US 2010/01201 10 A1 May 13, 2010 wo 02/072758 9/2002 wo 2010/019696 2/2010 Related US. Application Data OTHER PUBLICATIONS Walker & Croteau, “Molecular Cloning of a 10-deacetylbaccatin (63) Continuation of application No. 10/097,559, ?led on III-10-O-acetyl Transferase cDNA from Taxus and Functional Mar. 8, 2002, now Pat. No. 7,405,057. Expression in Escherichia coli,” Jan. 18, 2000, 97(2):583-587. Provisional application No. 60/274,421, ?led on Mar. Back et al., “Expression of a Plant Sesquiterpene Cyclase Gene in (60) Escherichia coli,” Archives in Biochemistry and Biophysics (1994) 9, 2001, provisional application No. 60/275,597, ?led 315:527-532. on Mar. 13, 2001. Mandujano-Chavez et al., “Differential Induction of Sesquiterpene Metabolism in Tobacco Cell Suspension Cultures by Methyl (51) Int. Cl. Jasmonate and Fungal Elicitor,” Archives in Biochemistry and Bio C12N1/00 (2006.01) physics (2000) 381:285-294. C12N1/06 (2006.01) Ralston et al.. “Cloning. Heterologous Expression, and Functional C12N 5/00 (2006.01) Characterization of 5-epi-arist010chene-1,3-Dihydr0xylase from Tobacco (Nicotiana tobacum), ” Archives in Biochemistry and Bio C12N 5/07 (2010.01) physics (2001) 393:222-235. (52) US. Cl. 435/69.1; 435/419; 435/348; 435/252.1; Zook et al., “Characterization of Novel Sesquiterpene Biosynthesis 435/252.2 in Tobacco Expressing Fungal Sesquiterpenoid Synthase,” Plant Field of Classi?cation Search ...................... .. None Physiology (1996) 112:311-318. (58) Akiyoshi-Shibata et al., “Further Oxidation of Hydroxycalcidiol by See application ?le for complete search history. Calcidiol 24-Hydr0xylase. A Study with the Mature Enzyme Expressed in Escherichia coli,” Eur. J. Biochem. (1994) 224:335 (56) References Cited 343. Back & Chappell, “Cloning and Bacterial Expression of a U.S. PATENT DOCUMENTS Sesquiterpene Cyclase from Hyoscyamus muticus and its Molecular Comparison to Related Terpene Cyclases,” J. Biol. Chem. (1995) 5,589,619 A 12/1996 Chappellet a1. ............ .. 800/205 5,672,487 A 9/1997 Schweden et a1. 270:7375-7381. 5,741,674 A 4/1998 Schweden et a1. Back & Chappell, “Identifying Functional Domains within Terpene 5,766,911 A 6/1998 Koike et al. ................. .. 435/193 Cyclases Using a Domain-Swapping Strategy,” Proc. Natl. Acad. Sci. 5,824,774 A 10/1998 Chappellet a1. USA. (1996) 93:6841-6845. 5,981,843 A 11/1999 Chappellet a1. ............ .. 800/301 5,994,114 A 11/1999 Croteau et a1. .............. .. 435/232 (Continued) 6,072,045 A 6/2000 Chappellet a1. Primary Examiner * Medina A Ibrahim 6,100,451 A 8/2000 Chappellet a1. ............ .. 800/298 (74) Attorney, Agent, 0rFirm * McKenna Long &Aldridge 6,117,649 A 9/2000 Bellamine et a1. 6,194,185 B1 2/2001 Croteau et a1. LLP; Stephanie Seidman 6,331,660 B1 12/2001 Chomet etal. (57) ABSTRACT 6,368,837 B1 4/2002 Gatenby et a1. 6,468,772 B1 10/2002 Chappellet a1. ............ .. 435/183 The invention features isolated cytochrome P450 polypep 6,495,354 B2 12/2002 Chappellet a1. 435/183 tides and nucleic acid molecules, as well as expression vec 6,531,303 B1 3/2003 Millis et a1. 435/155 tors and transgenic plants containing these molecules. In 6,559,297 B2 5/2003 Chappellet a1. .. 536/231 addition, the invention features uses of such molecules in 6,569,656 B2 5/2003 Chappellet a1. 435/183 methods of increasing the level of resistance against a disease 6,645,762 B2 11/2003 Chappellet a1. 435/325 6,689,593 B2 2/2004 Millis et a1. 435/155 caused by a plant pathogen in a transgenic plant, in methods 6,890,752 B2 5/2005 Chappellet a1. 435/325 for producing altered compounds, for example, hydroxylated 7,186,891 B1 3/2007 Chappellet a1. 800/298 compounds, and in methods of producing isoprenoid com 7,405,057 B2* 7/2008 Chappellet a1. .. 435/69.1 pounds. 7,442,785 B2 10/2008 Chappellet a1. .. 536/236 7,622,614 B2 11/2009 Julien et a1. ................. .. 568/327 20 Claims, 11 Drawing Sheets US 8,263,362 B2 Page 2 OTHER PUBLICATIONS Hutvagner et al., “Isolation and Sequence Analysis of a cDNA and a Back et al., “Cloning and Bacterial Expression of Sesquiterpene Related Gene for Cytochrome P450 Proteins from Solanum Cyclase, a Key Branch Point Enzyme for the Synthesis of chacoense,” Gene (1998) 188:247-252. Sesquiterpenoid Phytoalexin Capsidiol in UV-Challenged Leaves of Keller et al., “Sesquiterpene Cyclase Is not a Determining Factor for Capsicum annuum. ” Plant Cell Physiol. (1998) 39:899-904. Elicitor- and Pathogen-Induced Capsidiol Accumulation in Beckman et al., “Human 25-Hydroxyvitamin D3 -24-Hydroxylase, a Tobacco,” Planta (1998) 205:467-476. Multicatalytic Enzyme,” Biochemistry (1996) 35:8465-8472. Lupien et al., “Regiospeci?c Cytochrome P450 Limonene Boddupalli et al., “Fatty Acid Monooxygenation by P450BM-3: Hydroxylases from Mint (Mentha) Species: cDNA Isolation, Char Product Identi?cation and Proposed Mechanisms for the Sequential acterization, and Functional Expression of (—)-4S-Limonene-3 Hydroxylation Reactions,”Arch. Biochem. Biophys. (1992) 292:20 Hydroxylase and (—)-4S-Limonene-6-Hydroxylase.” Arch. 28. Biochem. Biophys. (1999) 368:181-192. Chappell et al., “Accumulation of Capsidiol in Tobacco Cell Cultures Mandujano-Chavez et al., “Differential Induction of Sesquiterpene Treated with Fungal Elicitor,” Phytochemisz (1987) 26:2259-2260. Chappell & Nable, “Induction of Sesquiterpenoid Biosynthesis in Metabolism in Tobacco Cell Suspension Cultures by Methyl Tobacco Cell Suspension Cultures by Fungal Elicitor,” Plant Physiol. Jasmonate and Fungal Elicitor,” Arch. Biochem. Biophys. (2000) (1987) 85:469-473. 381:285-294. Chappell, “Biochemistry and Molecular Biology of the Isoprenoid Mathis et al., “Pre-Steady-State Study of Recombinant Biosynthetic Pathway in Plants,” Annu. Rev. Plant Physiol. Plant Sesquiterpene Cyclases,” Biochemistry (1997) 36:8340-8348. Mol. Biol. (1995) 46:521-547. Maughan et al., “Expression of CYP71B7, a Cytochrome P450 Chapple, “Molecular-Genetic Analysis of Plant Cytochrome P450 Expressed Sequence Tag from Arabidopsis thaliana,” Arch. Dependent Monooxygenases.” Annu., Rev. Plant Physiol. Plant Mol. Biochem. Biophys. (1997) 341:104-111. Biol. (1998) 49:311-343. Milet et al., “Capsidiol and Ethylene Production by Tobacco Cells in Clark et al., “Spatially Distinct Expression of Two New Cytochrome Response to Cryptogein, an Elicitor from Phytophthora cryptogea,” P450s in Leaves of Nepeta racemosa: Identi?cation of a Trichome Phytochemistry (1991) 30:2171-2173. Speci?c Isoform,” Plant Mol. Biol. (1997) 33:875-885. Miller, “Structure of Genes Encoding Steriodogenic Enzymes,” J. Coolbaugh et al., “Studies on the Speci?city and Site ofAction ofa Steroid. Biochem. (1987) 27:759-766. alpha-Cyclopropyl-alpha-[p-Methoxphenyl]-5-Pyrimidine Methyl Molot et al., “Relations Between Capsidiol Concentration, Speed of Alcohol Amcymidol), a Plant Growth Regulator,” Plant Physiol. Fungal Invasion and Level of Induced Resistance in Cultivars of (1978) 62:571-576. Pepper (Capsicum annuum) Susceptible or Resistant to Cooper & Porter, “Mutagenicity of Nitrosamines in Phytophthora capsici,” Physiol. Plant Pathol. (1981) 18:379-389. Methyltransferase-De?cient Strains of Salmonella typhimurium Nedelkina et al., “Novel Characteristics and Regulation of a Diver Coexpressing Human Cytochrome P450 2E1 and Reductase,” Mutat. Res. (2000) 454:45-52. gent Cinnamate 4-Hydroxylase (CYP3A15) from French Bean: Diener et al., “Sterol Methyltransferase 1 Controls the Level of Cho Engineering Expression in Yeast,” Plant Mol.
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