Cytochrome P450-Dependent Metabolism of Oxylipins in Tomato. Cloning and Expression of Allene Oxide Synthase and Fatty Acid Hydroperoxide Lyase

Howe, Gregg A.; Lee, Gyu In; Itoh, Aya; Li, Lei; DeRocher, Amy E.

doi:10.1104/pp.123.2.711

Cited by 227 publications

(191 citation statements)

References 69 publications

Supporting

Mentioning

175

Contrasting

Unclassified

Order By: Relevance

“…The activity of several CYP74-enzymes has previously been described to be located in plastids [3]. Analysis of amino acid sequences of different cloned members of the CYP74 family revealed the presence of plastidic transit peptides [ 25,26,27]. The deduced amino acid sequence of AsDES lacks such a typical transit peptide.…”

Section: Isolation Of a Cyp74-enzyme From Garlicmentioning

confidence: 97%

Lipid metabolism in arbuscular mycorrhizal roots of Medicago truncatula

Stumpe¹,

Carsjens²,

Stenzel

et al. 2005

Phytochemistry

117

View full text Add to dashboard Cite

Section: Isolation Of a Cyp74-enzyme From Garlicmentioning

confidence: 97%

Lipid metabolism in arbuscular mycorrhizal roots of Medicago truncatula

Stumpe¹,

Carsjens²,

Stenzel

et al. 2005

Phytochemistry

117

View full text Add to dashboard Cite

“…Afterwards adapters were ligated to the full sequence at both 3 and 5 ends, and RACE-PCR was performed with Hifi-Taq polymerase (KAPA Biosystems, Boston, United States). First, degenerated primers were designed based on the conserved sequences of four HPLs [23][24][25][26]. After a first successful partial amplification of the BvHPL gene, four gene specific primers were designed for further amplifications (Forward R1: 5 -CCAAGTTCACGGGGATAGGAGAGTTGGTG-3 ; R2: 5 -GTGTAATTTAGCCTATCCCCTATCGG-3 ; Upstream R3: 5 -CCAAGTTCA-CGGGGATAGGAGAGTTGGTG-3 ; R4: 5 -AAGGTTAAAACCCCCTGTACCACTTC-3 ).…”

Section: Race-pcr For Bvhpl Genementioning

confidence: 99%

Optimization and scaling up of a biotechnological synthesis of natural green leaf volatiles using Beta vulgaris hydroperoxide lyase

Gigot

Ongena

Fauconnier

et al. 2012

Process Biochemistry

View full text Add to dashboard Cite

a b s t r a c tFollowing a promising preliminary study concerning sugar beet leaves valorization and to fulfill the high demand in natural C 6 -aldehydes flavors, an efficient biocatalytic reaction was developed to synthesize (2E)-hexenal at large scale. As major product of the lipoxygenase enzyme, 13-HPOT was converted by sugar beet hydroperoxide lyase extracted from leaves or expressed by recombinant Escherichia coli strains. With the adaptation of a fed-batch substrate addition and a continuous extraction of volatiles, 3.46 mM and 1.37 mM of C 6 -aldehydes were produced with the native hydroperoxide lyase at respectively 2 and 100 L scale. Furthermore, higher molar productivity of green leaf volatiles was reached with recombinant hydroperoxide lyase (5.5 mM at 2 L scale) while no others side products from the lipoxygenase pathway were formed. Once purified, these natural aromas are suitable for food and beverage preparations, perfumes or cosmetic products.

show abstract

“…This contrasts with known pathways in which lipoxygenase products are converted by the thiolate-ligated cytochrome P450s. To probe the significance of the coral fusion, reactions of the coral AOS domain with a pseudosubstrate of catalases have been examined.The AOS from coral is not related in sequence (6) to the allene oxide synthases that are abundant in plants (5,(10)(11)(12). Allene oxide synthase activity in plants is carried out by cytochrome P450 enzymes in the CYP74 group of atypical P450s (2).…”

mentioning

confidence: 99%

Role of Radical Formation at Tyrosine 193 in the Allene Oxide Synthase Domain of a Lipoxygenase−AOS Fusion Protein from Coral

Katsir

Seavy

et al. 2003

Biochemistry

View full text Add to dashboard Cite

Coral allene oxide synthase (cAOS), a fusion protein with 8R-lipoxygenase in Plexaura homomalla, is a hemoprotein with sequence similarity to catalases. cAOS reacts rapidly with the oxidant peracetic acid to form heme compound I and intermediate II. Concomitantly, an electron paramagnetic resonance (EPR) signal with tyrosyl radical-like features, centered at a g-value of 2.004-2.005, is formed. The radical is identified as tyrosyl by changes in EPR spectra when deuterated tyrosine is incorporated in cAOS. The radical location in cAOS is determined by mutagenesis of Y193 and Y209. Upon oxidation, native cAOS and mutant Y209F exhibit the same radical spectrum, but no significant tyrosine radical forms in mutant Y193H, implicating Y193 as the radical site in native cAOS. Estimates of the side chain torsion angles for the radical at Y193, based on the β-proton isotropic EPR hyperfine splitting, A iso , are θ 1 = 21 to 30° and θ 2 = −99 to −90°. The results show that cAOS can cleave nonsubstrate hydroperoxides by a heterolytic path, although a homolytic course is likely taken in converting the normal substrate, 8R-hydroperoxyeicosatetraenoic acid (8R-HpETE), to product. Coral AOS achieves specificity for the allene oxide formed by selection of the homolytic pathway normally, while it inactivates by the heterolytic path with nonoptimal substrates. Accordingly, with the nonoptimal substrate, 13R-hydroperoxyoctadecadienoic acid (13R-HpODE), mutant Y193H is inactivated after turning over significantly fewer substrate molecules than required to inactivate native cAOS or the Y209F mutant because it cannot absorb oxidizing equivalents by forming a radical at Y193. Polyunsaturated fatty acids (PUFAs) 1 are rapidly converted to cell signaling molecules in multistep syntheses involving an initial lipoxygenase or prostaglandin synthase step, followed by further enzymatic conversions (1-3). These multistep syntheses are regulated by localization of the enzymes in subcellular compartments and by protein-protein associations. Examples include synthesis of leukotriene C 4 at the nuclear envelope of leukocytes (4) and synthesis of jasmonic acid by steps which are thought to begin in chloroplasts and to be completed in peroxisomes of green plant cells (5). A fusion protein identified in the coral Plexaura homomalla represents a third mode of regulation, in which an N-terminal catalaselike domain is fused to an 8R-lipoxygenase domain (6). The N-terminal domain (44 kDa) has allene oxide synthase (cAOS) activity with the 8R-hydroperoxide of arachidonic acid (8R- † This research was supported by National Institutes of Health Grant GM36232 (to B.J.G.).* To whom correspondence should be addressed. Phone: (850) 644-8547. Fax: (850) 644-8547. E-mail: gaffney@bio.fsu.edu.. ‡ These authors contributed equally.1 Abbreviations: cAOS, coral allene oxide synthase; BLC, bovine liver catalase; BSA, bovine serum albumin; CYP, cytochrome P450; DES, divinyl ether synthase; EPR, electron paramagnetic resonance; 8R-HpETE, 8R-hydroperoxy-9E,11Z-ei...

show abstract

Cytochrome P450-Dependent Metabolism of Oxylipins in Tomato. Cloning and Expression of Allene Oxide Synthase and Fatty Acid Hydroperoxide Lyase

Cited by 227 publications

References 69 publications

Lipid metabolism in arbuscular mycorrhizal roots of Medicago truncatula

Lipid metabolism in arbuscular mycorrhizal roots of Medicago truncatula

Optimization and scaling up of a biotechnological synthesis of natural green leaf volatiles using Beta vulgaris hydroperoxide lyase

Role of Radical Formation at Tyrosine 193 in the Allene Oxide Synthase Domain of a Lipoxygenase−AOS Fusion Protein from Coral

Contact Info

Product

Resources

About