Structure of 3,4-Dihydroxy-2-butanone 4-Phosphate Synthase from Methanococcus jannaschii in Complex with Divalent Metal Ions and the Substrate Ribulose 5-Phosphate: implications for the catalytic mechanism

Skeletal rearrangements of carbohydrates are crucial for many biosynthetic pathways. In riboflavin biosynthesis ribulose 5-phosphate is converted into 3,4-dihydroxy-2-butanone 4-phosphate while its C4 atom is released as formate in a sequence of metal-dependent reactions. Here, we present the crystal structure of Methanococcus jannaschii 3,4-dihydroxy-2-butanone 4-phosphate synthase in complex with the substrate ribulose 5-phosphate at a dimetal center presumably consisting of non-catalytic zinc and calcium ions at 1.7-Å resolution. The carbonyl group (O2) and two out of three free hydroxyl groups (OH3 and OH4) of the substrate are metal-coordinated. We correlate previous mutational studies on this enzyme with the present structural results. Residues of the first coordination sphere involved in metal binding are indispensable for catalytic activity. Only Glu-185 of the second coordination sphere cannot be replaced without complete loss of activity. It contacts the C3 hydrogen atom directly and probably initiates enediol formation in concert with both metal ions to start the reaction sequence. Mechanistic similarities to Rubisco acting on the similar substrate ribulose 1,5-diphosphate in carbon dioxide fixation as well as other carbohydrate (reducto-) isomerases are discussed

Study on the Essential Oil of Aerial and Sub-Aerial Parts of Cymbopogon Flexuosus (Nees Ex Steud) Wats.

Cymbopogon flexuosus (Nees ex Steud) Wats commonly known as, East Indian lemongrass a widely grown essential oil plant in the world belongs to the family Poaceae and comprise of 140 species worldwide, found abundantly in tropics and sub-tropic regions of Asia, Africa and America. In India, 45 species are recorded of which the economic importance is C.winterianus, C.flexuosus, C.martinii, C.nardus, C.citratus, C.pendulus, C.jwarancusa and C.khasianus. Aerial and sub-aerial parts of C. flexuosus collected from Himavath Gopala hills, Karnataka, India, were subjected to hydrodistillation for extraction of essential oil. GC and GC-MS analysis were performed to know the chemical composition of the oil. Among the 39 compounds identified in aerial parts of the plant the major compounds were citral (64.98%), 1,7-octadien-3-ol (10.97%), dimethyl oxatricyclo nonanone (9.44%), nerol (2.85%), verbenol (1.77%) and caryophyllene oxide (0.71%). In sub-aerial parts of the plant 33 compounds were identified. The analysis of sub-aerial parts showed a different chemical profile compared to aerial part and possessed citral as the major compound of upto 30.47%. Other compounds in sub-aerial part are Eudesmol (17.82%), Elemol (14.16%), dihydro isopropyl methyl azulene (11.08%), .-cadinene (1.88%), junipene (1.36%), hydroxyalloaromadendrene, juniper camphor (1.12%) and elemene (1.04%)

Generation of Molecular Complexity from Cyclooctatetraene: Preparation of Aminobicyclo[5.1.0]octitols

A series of eight stereoisomeric N-(tetrahydroxy bicyclo-[5.1.0]oct-2S*-yl)phthalimides were prepared in one to four steps from N-(bicyclo[5.1.0]octa-3,5-dien-2-yl)phthalimide (±)-7, which is readily available from cyclooctatetraene (62 % yield). The structural assignments of the stereoisomers were established by 1H NMR spectral data as well as X-ray crystal structures for certain members. The outcomes of several epoxydiol hydrolyses, particularly ring contraction and enlargement, are of note. The isomeric phthalimides as well as the free amines did not exhibit β-glucosidase inhibitory activity at a concentration of less than 100 μM

Metabolites of 2,3-diketogulonate delay peroxidase action and induce non-enzymic H2O2 generation : Potential roles in the plant cell wall

A proportion of the plant's L-ascorbate (vitamin C) occurs in the apoplast, where it and its metabolitesmay act as pro-oxidants and anti-oxidants. One ascorbate metabolite is 2,3-dilcetogulonate (DKG), preparations of which can non-enzymically generate H2O2 and delay peroxidase action on aromatic substrates. As DKG itself generates several by-products, we characterised these and their ability to generate H2O2 and delay peroxidase action. DKG preparations rapidly produced a by-product, compound (1), with lambda(max) 271 and 251 nm at neutral and acidic pH respectively. On HPLC, (1) co-eluted with the major H2O2-generating and peroxidase-delaying principle. Compound (1) was slowly destroyed by ascorbate oxidase, and was less stable at pH 6 than at pH 1. Electrophoresis of an HPLC-enriched preparation of (1) suggested a strongly acidic (pK(a) approximate to 2.3) compound. Mass spectrometry suggested that un-ionised (1) has the formula C6H6O5, i.e. it is a reduction product of DKG (C6H8O7). In conclusion, compound (1) is the major H2O2-generating, peroxidase-delaying principle formed non-enzymically from DKG in the pathway ascorbate -> dehydroascorbic acid -> DKG -> (1). We hypothesise that (1) generates apoplastic H2O2 (and consequently hydroxyl radicals) and delays cell-wall crosslinking - both these effects favouring wall loosening, and possibly playing a role in pathogen defence. (C) 2017 The Authors. Published by Elsevier Inc.Peer reviewe

Aromatic Characterisation of Malvasia Odorosissima Grapevines and Comparison with Malvasia di Candia Aromatica

Malvasia odorosissima is an aromatic grapevine (Vitis vinifera L., 1753) variety that is often confused with Malvasia di Candia aromatica (Vitis vinifera L., 1753), despite the genetic information now available on the pedigree and genetic relationships linking the two varieties. In an effort to offer a contribution to fill this gap, also from an aromatic point of view, free and glycosylated aroma compounds were determined using the SPE extraction method, followed by GC-MS analysis in two consecutive vintages. The results have for the first time provided the aromatic characterisation of Malvasia odorosissima. Geraniol and its derivatives were the most abundant set of volatiles. In contrast to Malvasia di Candia aromatica and the other aromatic varieties, Malvasia odorosissima showed a very small amount of glycosylated volatiles, thus expressing its aromatic potential almost completely. The abundance of free terpenoids in the aromatic profile of Malvasia odorosissima, even higher than in Malvasia di Candia aromatica, is a main feature for the oenological exploitation of this variety, which is on the brink of extinction. In addition, the presence of rose oxides, found solely in Malvasia odorosissima, renders its aromatic profile more similar to that of White Muscat. This result is consistent with the parent-offspring relationship linking the two varieties that was recently ascertained