Coral allene oxide synthase (AOS), a hemoprotein with weak sequence homology to catalase, is the N-terminal domain of a naturally occurring fusion protein with an 8R-lipoxygenase. AOS converts 8R-hydroperoxyeicosatetraenoic acid to the corresponding allene oxide. The UV-visible absorption and magnetic circular dichroism spectra of ferric AOS and of its cyanide and azide complexes, and the electron paramagnetic resonance spectra of native AOS (high-spin, g ) 6.56, 5.22, 2.00) and of its cyanide adduct (low-spin, g ) 2.86, 2.24, 1.60) closely resemble the corresponding spectra of bovine liver catalase (BLC). These results provide strong evidence for tyrosinate ligation to the heme iron of AOS as has been established for catalases. On the other hand, the positive circular dichroism bands in the Soret region for all three derivatives of ferric AOS are almost the mirror image of those in catalase. In addition, the cyanide affinity of native AOS (K d ) 10 mM at pH 7) is about 3 orders of magnitude lower than that of BLC. Thus, while these results conclusively support a common tyrosinate-ligated heme in AOS as in catalase, significant differences exist in the interaction between their respective heme prosthetic groups and protein environments, and in the access of small molecules to the heme iron.Interest in the enzymology underlying the high prostaglandin content of the Caribbean sea whip coral, Plexaura homomalla, has been the impetus for several biochemical investigations. Arachidonic acid is metabolized in extracts of P. homomalla and related corals by a lipoxygenase (LOX) 1 pathway that appeared initially to relate to the production of cyclopentenone prostaglandins (1, 2). An 8R-lipoxygenase converts arachidonic acid to 8R-hydroperoxyeicosatetraenoic acid (8R-HPETE), and the fatty acid hydroperoxide is then enzymatically dehydrated to an allene oxide, an epoxide with a propensity to cyclize to cyclopentenone derivatives. Brash et al., in 1997, isolated the cDNA of an 8R-lipoxygenase from P. homomalla and found it to occur with an extra sequence in the open reading frame that encodes the allene oxide synthase (AOS) (3). This natural fusion protein in coral consists of a LOX C-terminal domain (79 kDa) and an AOS N-terminal domain (43 kDa). The truncated (AOS) construct catalyzes an identical reaction to the AOS domain in the fusion protein, converting 8R-HPETE solely to the allene oxide. The truncated construct exhibits very high turnover number (1400/s) (4), although, due to the weaker expression of the fusion protein in E. coli, the comparable values for the fusion protein remain to be determined. 2 Although allene oxide has not proven to be an intermediate in the prostaglandin synthesis pathway (5), it may be linked to prostanoid type products such as clavulones and punaglandins of other corals (2,6,7). Allene oxides are very unstable epoxides; they readily hydrolyze into ketols, cyclopentenones, and other rearrangement products (8-10).The production of allene oxides from fatty acid hydroperoxides also ...

Xylella fastidiosa is the causal agent of Pierce's disease in grapevines. The mechanisms of pathogenicity are largely due to occlusion of xylem vessels by aggregation of X. fastidiosa and biofilm formation. Xylella fastidiosa was subjected to xylem fluids with varying chemistries to examine the effects of nutritional components on bacterial growth in vitro. The exposure of X. fastidiosa to xylem fluids collected from different Vitis genotypes resulted in highly significant differences in both planktonic growth and biofilm formation. Planktonic growth of X. fastidiosa in Vitis xylem fluid was correlated to the concentration of citric acid, amino acids (glutamic acid, glutamine, histidine, valine, methionine, isoleucine and phenylalanine) and inorganic ions (copper, magnesium, phosphorus and zinc). Biofilm formation was correlated to many amino acids at 1 h of incubation. Xylem fluid from Vitis rotundifolia cv. Noble (fluid that supported low planktonic growth) was supplemented with the compounds that were correlated above to levels found in Vitis champinii cv. Ramsey (fluid that supported high planktonic growth) to determine the direct impact of xylem constituents on the growth characteristics of X. fastidiosa. Augmentation of fluid from Noble with the amino acids listed above, citric acid, calcium and magnesium resulted in increased planktonic growth and aggregation.