Isolation and characterization of rat intestinal bacteria involved in biotransformation of (−)-epigallocatechin

Two intestinal bacterial strains MT4s-5 and MT42 involved in the degradation of (?)-epigallocatechin (EGC) were isolated from rat feces. Strain MT4s-5 was tentatively identified as Adlercreutzia equolifaciens. This strain converted EGC into not only 1-(3, 4, 5-trihydroxyphenyl)-3-(2, 4, 6-trihydroxyphenyl)propan-2-ol (1), but also 1-(3, 5-dihydroxyphenyl)-3-(2, 4, 6-trihydroxyphenyl)propan-2-ol (2), and 4′-dehydroxylated EGC (7). Type strain (JCM 9979) of Eggerthella lenta was also found to convert EGC into 1. Strain MT42 was identified as Flavonifractor plautii and converted 1 into 4-hydroxy-5-(3, 4, 5-trihydroxyphenyl)valeric acid (3) and 5-(3, 4, 5-trihydroxyphenyl)-γ-valerolactone (4) simultaneously. Strain MT42 also converted 2 into 4-hydroxy-5-(3, 5-dihydroxyphenyl)valeric acid (5), and 5-(3, 5-dihydroxyphenyl)-γ-valerolactone (6). Furthermore, F. plautii strains ATCC 29863 and ATCC 49531 were found to catalyze the same reactions as strain MT42. Interestingly, formation of 2 from EGC by strain MT4s-5 occurred rapidly in the presence of hydrogen supplied by syntrophic bacteria. Strain JCM 9979 also formed 2 in the presence of the hydrogen or formate. Strain MT4s-5 converted 1, 3, and 4 to 2, 5, and 6, respectively, and the conversion was stimulated by hydrogen, whereas strain JCM 9979 could catalyze the conversion only in the presence of hydrogen or formate. On the basis of the above results together with previous reports, the principal metabolic pathway of EGC and EGCg by catechin-degrading bacteria in gut tract is proposed.     

Development of SCAR marker for sex identification in dioecious Garcinia gummi-gutta

Key message

We have developed sex-specific SCAR marker for the identification of dioecious Garcinia gummi - gutta (L.), which is useful for the selection of G. gummi - gutta at seedling stage and for plantation programmes.

Abstract

Garcinia gummi-gutta (L.) Robs. is a dioecious fruit yielding tree, which is naturally distributed as well as cultivated in the orchards in Western Ghat regions of India. A sex-linked DNA fragment was identified in Garcinia gummi-gutta (L.) Robs. by screening 150 randomly amplified polymorphic DNA primers and only one of them (OPBD20) showed different amplification band pattern associated with sex type. This sex-linked fragment was converted into male-specific sequence-characterized amplified region (SCAR) marker, CAM-566. The primers deigned in this study (OPBD20F and OPBD20R) correctly differentiated 12 male and 12 female plants at high annealing temperatures. Thus, a 556-bp band was amplified in male samples but not in female ones. Nevertheless, it should be noted that the fragments from both sexes were amplified at relatively low annealing temperatures. Additionally, the developed SCAR marker successfully identified the sexes of ten sex-unknown samples. Therefore, it can be used as an effective, convenient and reliable tool for sex determination in such dioecious species.     

Synthesis and Reactions of Some 2-Aminobenzothiazoles

N-(2-Benzothiazolyl)- and N-(6-methoxy-2-benzothiazolyl)cyanoacetamides 4, 5 resulted in the reaction of 2-aminobenzothiazole 1 or its 6-methoxy derivative 2 with 1-cyanoacetyl-3,5-dimethylpyrazole 3. Both cyanoacetylamides 4 and 5 have been transformed into the corresponding 2-oxo-2H-pyrimido[2,1-b]-benzothiazole-3-carbonitrile 8 and its 8-methoxy derivative 9 by reaction with triethyl orthoformate, followed by cyclization.     

First laccase in green algae: purification and characterization of an extracellular phenol oxidase from Tetracystis aeria

Main conclusion

A green algal phenol oxidase was firstly purified, confirmed to be a laccase, and a hetero-oligomeric quaternary structure is suggested. The operation of a laccase-mediator system is firstly described in algae. Laccases (EC 1.10.3.2) catalyze the oxidation of a multitude of aromatic substrates. They are well known in higher plants and fungi, while their presence in green algae appears uncertain. Extracellular laccase-like enzyme activity has previously been described in culture supernatants of the green soil alga Tetracystis aeria [Otto et al. in Arch Microbiol 192:759–768, (2010)]. As reported herein, the T. aeria enzyme was purified 120-fold by employing a combination of anion exchange and size exclusion chromatography. The purified enzyme was confirmed to be a laccase according to its substrate specificity. It oxidizes 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), syringaldazine, and 2,6-dimethoxyphenol (pH optima of pH ≤2.5, 7.0, and 6.5; K _m values of 28.8, 40.5, and 1,830 µM; respectively), but not l-tyrosine or Fe²⁺. ABTS is by far the most efficient substrate. Two polypeptides, A (~110 kDa) and B (71 kDa), were co-purified by the applied procedure, both being highly N-glycosylated (≥~53 and ≥27 %, respectively). As suggested by various gel electrophoretic analyses, the native enzyme (apparent molecular mass of ~220 kDa) most probably is a hetero-oligomer with the composition AB ₂ , wherein A is the catalytic subunit and B forms a disulfide-linked homo-dimer B ₂ . The decolorization of anthraquinone (Acid Blue 62 and Remazol Brilliant Blue R) and diazo dyes (Reactive Black 5) was studied in the presence of redox-mediating compounds (ABTS and syringaldehyde), demonstrating the operation of the laccase-mediator system in algae for the first time. Thus, laccases from green algae may participate in the biotransformation of a wide spectrum of natural and xenobiotic compounds.     

Computational comparison of the kinetic stabilities of diamino- and diamidocarbenes in the 1,2-H shift reaction

In this study, we performed several DFT, MP2, and BD(T) calculations on the 1,2-H shift reactions of two diaminocarbenes (1, 2) and a diamidocarbene (3) using the Gaussian 09 program. In Gaussian 09, the BD(T) method keyword requests a Brueckner doubles calculation including a perturbative triples contribution. Although N-heterocyclic carbenes (NHC) are typically known for their exceptional σ-donor abilities, recent studies have indicated that π-interactions also play a role in the bonding between NHCs and transition metals or BX₃ (X = H, OH, NH₂, CH₃, CN, NC, F, Cl, and Br) (Nemcsok et al. Organomet 23:3640–3646, 2004, Esrafili. J Mol Model 18:2003–2011, 2012). In order to study the importance of π-interactions between carbenes and transition metals, Hobbs and co-workers (Hobbs et al. New J Chem 34:1295–1308, 2010) focused on the synthesis of NHCs with reduced-energy lowest unoccupied molecular orbitals. By introducing an oxalamide moiety into the heterocyclic backbone, they found the resulting carbene possessed higher electrophilicity than usual NHCs. According to our results, the N,N'-diamidocarbene should be more stable than the diaminocarbenes with respect to the 1,2-H shift reaction.

Driving and retarding forces in a chemical reaction

The reaction force F(ξ) is the negative gradient of the potential energy of a chemical process along the intrinsic reaction coordinate ξ. We extend the rigorous concept of F(ξ) to the “activation strain model” of Bickelhaupt et al., to formulate the “strain” force F _str(ξ) that retards a reaction and the “interaction” force F _int(ξ) that drives it. These are investigated for a group of Diels-Alder cycloadditions. The results fully support the interpretation of the minimum of F(ξ) as defining the beginning of the transition from deformed reactants to eventual products.     

Mono- and digalactosyldiacylglycerols: potent nitric oxide inhibitors from the marine microalga Nannochloropsis granulata

Chemical investigation of a marine microalga, Nannochloropsis granulata, led to the isolation of four digalactosyldiacylglycerols namely, (2S)-1-O-eicosapentaenoyl-2-O-palmitoyl-3-O-(β-d-galactopyranosyl-6-1α-d-galactopyranosyl)-glycerol (1), (2S)-1-O-eicosapentaenoyl-2-O-palmitoleoyl-3-O-(β-d-galactopyranosyl-6-1α-d-galactopyranosyl)-glycerol (2), (2S)-1-O-eicosapentaenoyl-2-O-myristoyl-3-O-(β-d-galactopyranosyl-6-1α-d-galactopyranosyl)-glycerol (3), and (2S)-1,2-bis-O-eicosapentaenoyl-3-O-(β-d-galactopyranosyl-6-1α-d-galactopyranosyl)-glycerol (4), together with their monogalactosyl analogs (5–8). Among the isolated galactolipids 2 and 3 were new natural products. Complete stereochemistry of 1, 4, 5, 7, and 8 was determined for the first time by both spectroscopic techniques and classical degradation methods. Both mono- and digalactosyldiacylglycerols isolated from N. granulata possessed strong nitric oxide (NO) inhibitory activity against lipopolysaccharide-induced NO production in RAW264.7 macrophage cells through downregulation of inducible nitric oxide synthase expression indicating the possible use as anti-inflammatory agents.     

New species of hansteinia (acantaceae) from Colombia and Ecuador

New species of Hansteinia (Acanthacene) from Columbia and Equador, Brittonia 37: 203-208. 1985. – Two new species and a new variety, Hansteinia jamesonaii from Pichincha, Equador and H. Magdalensis and H. magdalenensis var. pilosa from the Sierra Nevada de Sta. Marta, are described, illustrated, and compared with H. charitopes their closest relative.     

Medium-β free-boundary equilibria of a quasi-isodynamic stellarator

Free-boundary MHD equilibria with magnetic surfaces in the vacuum region surrounding the plasma [E. Strumberger, Nucl. Fusion 37, 19 (1997); M. Drevlak, D. Monticello, and A. Reiman, Nucl. Fusion 45, 731 (2005)] are obtained for a quasi-isodynamic stellarator [A. A. Subbotin, M. I. Mikhailov, V. D. Shafranov et al., Nucl. Fusion 46, 921 (2006); M. I. Mikhailov, J. Nuhrenberg, and V. D. Shafranov, Plasma Phys. Rep. 35, 529 (2009)].     

Isolation and identification of an endophytic fungus Pezicula sp. in Forsythia viridissima and its secondary metabolites

In a survey of endophytic fungal biodiversity, an antimicrobial endophytic isolate zjwcf069 was obtained from twigs of Forsythia viridissima, Zhejiang Province, Southeast China. Zjwcf069 was then identified as Pezicula sp. through combination of morphological and phylogenetic analysis based on ITS-rDNA. Zjwcf069 here represented the first endophytic fungus in Pezicula isolated from host F. viridissima. From the fermentation broth, four compounds were obtained through silica gel column chromatography and Sephadex LH-20 under the guide of bioassay. Their structures were elucidated by spectroscopic analysis as mellein (1), ramulosin (2), butanedioic acid (3), and 4-methoxy-1(3H)-isobenzofuranone (4). Compound 4 here stood for the very first time as natural product from microbes. In vitro antifungal assay showed that compound 1 displayed growth inhibition against 9 plant pathogenic fungi, especially Botrytis cinerea and Fulvia fulva with EC₅₀ values below 50 μg/mL. Endophytic fungi in medicinal plants were good resources for bioactive secondary metabolites.     

New taxa and combinations in Siphonoglossa (Acanthaceae)

As presently known, the genusSiphonoglossa can be divided into two “subgenera,” one of which is here divided into two sections,Pentaloba andSiphonoglossa. Two new species are described from a locality in Durango, Mexico :S. durangensis in sect.Siphonoglossa andS. linearifolia in sect.Pentaloba. Three other species are transferred intoSiphonoglossa:S. canbyi from northeastern Mexico;S. buchii from Haiti and the Dominican Republic andS. incerta from southern Baja California, Mexico.     

Synthesis of 1,6-Diaryl-5,7-dioxo(dithio)imidazo[1,2-a] [1,3,5]triazines. A Novel Pathway for Imidazo[1,2-a] [1,3,5]triazine Systems Obtained via 1-(Imidazolin-2-yl)urea and Thiourea Derivatives

Novel 1,6-diaryl-5,7(1H)dioxo(dithio)-2,3-dihydroimidazo[1,2-a][1, 3, 5]triazines 8, and 9 were synthesized by cyclization of the respective 1-(imidazolin-2-yl)ureas 4 or thioureas 6 with phosgene or thiophosgene in the presence of bases. 1-Aryl-2-aminoimidazolines 1 reacting with arylisocyanates 2 or arylisothiocyanates 3 form a mixture of isomeric imidazolin-2-yl 4 and 6 and imidazolin-3-yl 5 and 7 urea or thiourea derivatives. Isomers 4 and 6 can be easily separated and used for the cyclization reaction. The structures of the main intermediates and the final target compounds were confirmed by ¹H-NMR spectral analysis. Discussion of the possible course of the reactions is also presented.     

New taxa and nomenclatural changes in arceuthobium (viscaceae)

The following new taxa are recognized inArceuthobium: subgeneraArceuthobium andVaginata; sectionsVaginata, Campylopoda, andMinuta; seriesCampylopoda,Rubra, andStricta; speciesA. apachecum,A. californicum,A. guatemalense,A. hondurense, andA. pini; new formae spécialesA. abietinum f. sp.concoloris,A. abietinum f. sp.magnificae. New combinations:A. abietinum (Engelm.) Hawksworth & Wiens, andA. microcarpum (Engelm.) Hawksworth & Wiens.     

N(4)-Tolyl-2-acetylpyridine thiosemicarbazones and their platinum(II,IV) and gold(III) complexes: cytotoxicity against human glioma cells and studies on the mode of action

Complexes [Au(2Ac4oT)Cl][AuCl₂] (1), [Au(H_py2Ac4mT)Cl₂]Cl·H₂O (2), [Au(H_py2Ac4pT)Cl₂]Cl (3), [Pt(H2Ac4oT)Cl]Cl (4), [Pt(2Ac4mT)Cl]·H₂O (5), [Pt(2Ac4pT)Cl] (6) and [Pt(L)Cl₂OH], L = 2Ac4mT (7), 2Ac4oT (8), 2Ac4pT (9) were prepared with N(4)-ortho- (H2Ac4oT), N(4)-meta- (H2Ac4mT) and N(4)-para- (H2Ac4pT) tolyl-2-acetylpyridine thiosemicarbazone. The cytotoxic activities of all compounds were assayed against U-87 and T-98 human malignant glioma cell lines. Upon coordination cytotoxicity improved in 2, 5 and 8. In general, the gold(III) complexes were more cytotoxic than those with platinum(II,IV). Several of these compounds proved to be more active than cisplatin and auranofin used as controls. The gold(III) complexes probably act by inhibiting the activity of thioredoxin reductase enzyme whereas the mode of action of the platinum(II,IV) complexes involves binding to DNA. Cells treated with the studied compounds presented morphological changes such as cell shrinkage and blebs formation, which indicate cell death by apoptosis induction.     

Characterization of the sugar-O-methyltransferase LobS1 in lobophorin biosynthesis

Lobophorins A (1) and B (2) belong to a large group of spirotetronate natural products with potent antibacterial and antitumor activities. The cloning of the lobophorin biosynthesis gene cluster from the deep-sea-derived Streptomyces sp. SCSIO 01127 identified a sugar-O-methyltransferase-encoding gene lobS1. The lobS1 inactivation mutant accumulated two new lobophorin analogs 3 and 4, different from 1 and 2 by lacking the 4-methyl group at the terminal l-digitoxose, respectively. Biochemical experiments verified that LobS1 was a SAM-dependent sugar-O-methyltransferase that required divalent metal ions for better activity. Antibacterial assays revealed compounds 3 and 4 were generally less potent than compounds 1 and 2. These findings suggest that the methylation on the terminal digitoxose by LobS1 tailors lobophorin biosynthesis and highlights the importance of this methylation for antibacterial potence.     

Revision of Plerandra (Araliaceae). I. A synopsis of the genus with an expanded circumscription and a new infrageneric classification

Phylogenetic studies have demonstrated that Schefflera, the largest genus of Araliaceae, is grossly polyphyletic, comprising five distinct clades within the family. In an effort to establish monophyletic genera among the elements that currently comprise Schefflera, the genus Plerandra is expanded to encompass all of the members of one of these clades. In this synoptical revision, a new infrageneric classification is presented (along with a key) in which six subgenera are recognized. Four of these subgenera are newly described (Plerandra subgenera Canacoschefflera, Costatae, Gabriellarum, and Veilloniorum) and a fifth represents a new combination (Plerandra subg. Dizygotheca). A total of 33 species (one with two subspecies) are accepted, one of which is newly described (P. veilloniorum), and 22 new combinations are made (P. actinostigma, P. baillonii, P. cabalionii, P. costata, P. crassipes, P. elegantissima, P. elongata, P. emiliana, P. gabriellae, P. leptophylla, P. nono, P. osyana, P. osyana subsp. toto, P. pachyphylla, P. pancheri, P. plerandroides, P. polydactylis, P. reginae, P. seemanniana, P. tannae, P. vanuatua, P. veitchii). Neotypes are provided for six accepted names and one heterotypic synonym, and lectotypes are designated for 13 accepted names and 16 heterotypic synonyms. For each accepted species, full synonymy is provided along with geographic range and notes.     

Orthologs and paralogs - we need to get it right

A response to Homologuephobia, by Gregory A Petsko, Genome Biology 2001 2:comment1002.1-1002.2, to An apology for orthologs - or brave new memes by Eugene V Koonin, Genome Biology 2001, 2:comment1005.1-1005.2, and to Can sequence determine function? by John A Gerlt and Patricia C Babbitt, Genome Biology 2000, 1:reviews0005.1-0005.10.     

Protonation,alkylation, addition and redox reactions of 16, 17 and 18 valence electron [Mo(L)(NO)('S4')] complexes [L = SPh,PMe3, NO; 'S4'2– = 1,2-Bis-(2-mercaptophenylthio)ethane(2-)]

In the quest for complexes modelling functional characteristics of metal sulfur oxidoreductases, a series of molybdenum nitrosyl complexes with sulfur-dominated coordination sphere was synthesized. Treatment of the 16, 17 and 18 valence electron (VE) complexes [Mo(L)(NO)('S₄')] (1–3) [L?=?SPh (1), PMe₃ (2), NO (3), 'S₄'^2–?=?1,2-bis-(2-mercaptophenylthio) ethane(2-)] with the Brönsted acid HBF₄ resulted in formation of different types of products. 1 and 3 were reversibly protonated at one thiolate atom of the 'S₄'^2– ligand;2, however, yielded the phosphonium salt [HPMe₃]BF₄ and the dinuclear [Mo(NO)('S₄')]₂. Alkylation of 1, 2 and 3 by Me₃OBF₄ or Et₃OBF₄ uniformly resulted in high yields of [Mo(L)(NO)(R-'S₄')]BF₄ complexes [L?=?SPh: R?=?Me (5), Et (6); L?=?PMe₃: R?=?Me (7); L?=?NO: R?=?Me (8), Et (9)] in which one thiolate atom of the 'S₄'^2– ligand had become alkylated; the NMR spectra of 5, 6, 8 and 9 indicated that only one out of four theoretically possible diastereoisomers had formed. 5 and 6 were characterized also by single-crystal X-ray structure analyses. A comparison of ν(NO) bands and redox potentials (cyclic voltammetry) of parent complexes and alkylated derivatives showed that alkylation leads to a decrease in electron density at the molybdenum center and to a positive shift in redox potentials. The 16 VE complex 1 could be reduced, also chemically, to give the corresponding 17 VE anion [1]^–, and inserted elemental sulfur into the Mo-SPh bond, forming the 18 VE phenylperthio complex [Mo(η²–SSPh)(NO)('S₄')] (11) which, upon reaction with PPh₃, gave SPPh₃ and regenerated the parent complex 1. These results are discussed with regard to the sequence of proton and electron transfer steps occurring in substrate conversions catalyzed by metal sulfur oxidoreductases.