Synthesis of p-nitrophenyl sulfated disaccharides with beta-D-(6-sulfo)-GlcNAc units using beta-N-acetylhexosaminidase from Aspergillus oryzae in a transglycosylation reaction

When α-d-GlcNAc-OC₆H₄NO₂ -p and β-d-(6-sulfo)-GlcNAc-OC₆H₄NO₂-p (2) were used as substrates, β-N-acetylhexosaminidase from Aspergillus oryzae transferred the β-d-(6-sulfo)-GlcNAc(unit from 2 to α-d-GlcNAc-OC₆H₄NO₂ -p to afford β-d-(6-sulfo)-GlcNAc-(1→4)-α-d-GlcNAc-OC₆H₄NO₂-p (3) in a yield of 94% based on the amount of donor, 2, added. β-d-(6-sulfo)-GlcNAc-(1→4)-α-d-Glc-OC₆H₄NO₂-p (4) was obtained with α-d-Glc-OC₆H₄NO₂ -p as acceptor in a similar manner. With a reaction mixture of 2 and β-d-GlcNAc-OC₆H₄NO₂-p (1) in a molar ratio of 6:1, the enzyme mediated the transfer of β-d-GlcNAc from 1 to 2, affording disaccharide β-d-GlcNAc-(1→4)-β-(6-sulfo)-d-GlcNAc-OC₆H₄NO₂-p (5) in a yield of 13% based on the amount of 1 added.     

A Novel Zeolite Coating for Protection of Concrete Sewers from Biological Sulfuric Acid Attack

Microbial growth inhibition and resistance to biological deterioration of concrete specimens coated with silver-loaded zeolite was evaluated by measuring the time course of bacterial growth, biological sulfur oxidation, and sulfate production using Acidithiobacillus thiooxidans as a corrosive agent. Live bacterial cells declined from an initial inoculum concentration of 1.1 × 10₄ cell ml_-1 to zero in 10 days, during which only 0.5–1% of the initial sulfur concentration of 10 g l_-1 was biologically oxidized, corresponding to sulfate production rates of 35–42 mg SO ₄ ^{2 ?} g ^{? 1} S ^{? 1} . Leaching coefficients of calcium and silicon in the specimens coated with silver-loaded zeolite of 1.6 × 10 ^{? 4} to 4.6 × 10 ^{? 2} cm ² d ^{? 1} respectively, were only 0.8% and 1% of the uncoated specimens.     

Studies Towards the Large Scale Chemical Synthesis of the Precursors of Ribonucleosides-3′,4′,5′,5″- 2H 4 and -2′,3′,4′,5′,5″- 2H 5

Abstract

A summary delineating the large scale synthetic studies to prepare labeled precursors of ribonucleosides-3′,4′,5′,5″- ²H ₄ and -2′,3′,4′,5′,5″- ²H ₅ from D-glucose is presented. The recycling of deuterium-labeled by-products has been devised to give a high overall yield of the intermediates and an expedient protocol has been elaborated for the conversion of 3-O-benzyl-α,β-D-allofuranose-3,4-d ₂ 6 to 1-O-methyl-3-O-benzyl-2-O-t-butyldimethylsilyl-α,β-D-ribofuranose-3,4,5,5′-d ₄ 16 (precursor of ribonucleosides-3′,4′,5′,5″- ²H ₄ ) or to 1-O-methyl-3,5-di-O-benzyl-α,β-D-ribofuranose-3,4,5,5′-d ₄ 18 (precursor of ribonucleosides-3′,4′,5′,5″- ²H ₄ ).     

Synthesis and conformational analysis of 1,2-cis fused bicyclic α-d-galactofuranosyl thiocarbamate from per-O-tert-butyldimethylsilyl-β-d-galactofuranosyl isothiocyanate

Per-O-tert-butyldimethylsilyl-α,β-d-galactofuranosyl isothiocyanate (4) was synthesized by the reaction of per-O-TBS-β-d-galactofuranose (1) with KSCN, promoted by TMSI. Upon O-desilylation (1,2-dideoxy-α-d-galactofuranoso)[1,2d]-1,3-oxazolidine-2-thione (6), the cis-fused bicyclic thiocarbamate was obtained as the only product. Conformational analysis, aided by molecular modelling, showed two stable twist forms (³T₄ and ⁴T_O) for the five-membered sugar ring in 6. In aqueous solution, the equilibrium favours the first conformation (3:1 ratio). On the other hand, this ratio decreases for less polar solvents.     

Regioselective Synthesis of Indazole N 1‐ and N 2‐(β‐d‐Ribonucleosides)

The regioselective synthesis of 4‐nitroindazole N ¹‐ and N ²‐(β‐d‐ribonucleosides) (8, 9, 1b and 2b) is described. The N ¹‐regioisomers are formed under thermodynamic control of the glycosylation reaction [fusion reaction or Silyl Hilbert‐Johnson glycosylation for 48 h (66%)], while the kinetic control (Silyl Hilbert‐Johnson glycosylation for 5 h) afforded only the N ²‐isomer (64%). The structures of the nucleosides 1b and 2b were assigned by single crystal X‐ray analyses. The 4‐amino‐N ¹‐(β‐d‐ribofuranosyl)‐1H‐indazole (3b) was obtained from the nitro nucleoside 1b by catalytic hydrogenation. Compound 3b shows fluorescence while the 4‐nitroindazole nucleosides 1b and 2b do not possess this property.     

Phosphoralaninate Pronucleotides of Pyrimidine Methylenecyclopropane Analogues of Nucleosides: Synthesis and Antiviral Activity

The Z- and E-thymine and cytosine pronucleotides 3d, 4d, 3e, and 4e of methylenecyclopropane nucleosides analogues were synthesized, evaluated for their antiviral activity against human cytomegalovirus (HCMV), herpes simplex virus 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), Epstein-Barr virus (EBV), human immunodeficiency virus type 1 (HSV-1), and hepatitis B virus (HBV) and their potency was compared with the parent compounds 1d, 2d, 1e, and 2e. Prodrugs 3d and 4d were obtained by phosphorylation of parent analogues 1d or 2d with reagent 8. A similar phosphorylation of N⁴-benzoylcytosine methylenecyclopropanes 9a and 9b gave intermediates 11a and 11b. Deprotection with hydrazine in pyridine–acetic acid gave pronucleotides 3e and 4e. The Z-cytosine analogue 3e was active against HCMV and EBV. The cytosine E-isomer 4e was moderately effective against EBV.     

Stereoselective microbial reduction of N-(4-(1-oxo-2-chloroacetyl ethyl) phenyl methane sulfonamide

Several microbial cultures were screened for the ability to catalyse the reduction of N-(4-(1-oxo-2-chloroacetyl ethyl) phenyl methane sulfonamide (1). The chiral intermediate (+)N-(4-(1-hydroxy-2-chloroethyl) phenyl methane sulfonamide (2) was prepared by the stereoselective microbial reduction of the parent ketone 1. Compound 2 is a potential chiral intermediate for synthesis of 4-(2-isopropylamino-1-hydroxyethyl) phenyl methanesulfonanilide (d-sotalol), a beta-receptor antagonist. Microorganisms from the genera Rhodococcus, Nocardia, and Hansenula reduced 1 to 2. A reaction yield of >50% and optical purities of >90% were obtained. The best strain (H.polymorpha ATCC 26012) effectively reduced compound 1 to compound 2 in 95% reaction yield and 99% optical purity. Compound 2 (8.2 g) was isolated from a 3-1 preparative batch in 68% overall yield. Isolated compound 2 had a specific rotation of +20° (CH₂Cl₂, C-1), an optical purity of 99.5%, and a chemical purity of 97% as analyzed by gas chromatography and HPLC. The nuclear magnetic resonance and mass spectra of compound 2 prepared by bioreduction and a standard chemical sample of 2 were virtually identical. Cell extracts of H. polymorpha in the presence of glucose dehydrogenase, glucose and nicotinamide adenine dinucleotide (NAD ⁺) catalyzed the reduction of 1 to 2 with 98% reaction yield and resulted in an optical purity of 99.4%. Correspondence to: R. N. Patel     

An Efficient Method for the Synthesis of β-d-Ribonucleosides Catalyzed by Metal Iodides

Abstract

Several β-d-ribonucleosides were synthesized in high yields under mild conditions by N-glycosylations of methyl 2,3,5-tri-O-benzoyl-β-d-ribofuranosyl carbonate (1) with trimethylsilylated nucleoside bases in acetonitrile using a catalytic amount of metal iodide such as SnI₂, SbI₃ or TeI₄. A deprotection of N⁶ -benzoyl group of coupling product took place to a considerable extent when N⁶ -benzoyl-N⁶ , N⁹ -bis(trimethylsilyl)adenine was employed as a nucleoside base using SnI₂ or SnCI₂ as a catalyst while it was minimized when SbI₃ or TeI₄ was used. Further, the N-glycosylation of 1 with 7-trimethylsilyltheophylline in the presence of a catalytic amount of metal iodide was more effectively achieved in nitrile solvents other than acetonitrile.

     

Polydeoxyaminohexopyranosylnucleosides. Synthesis of 1-(2,3,4-Trideoxy-3-nitro-β-D-erythro- and threo-hexopyranosyl)-uracils from Uridine

Abstract

The first synthesis of nitro-multideoxy-sugar containing nucleosides was achieved. 1-(4,6-O-Benzylidene-3-deoxy-3-nitro-β-D-glucopyranosyl)uracil (3) was converted in 75% yield into 1-(4,6-O-benzylidene-2,3-dideoxy-3-nitro-arabinohexopyranosyl)uracil (7) by acetylation followed by NaBH₄ reduction in methanol. De-O-benzylidenation with CF₃CO₂H afforded crystalline 1-(2,3-dideoxy-3-nitro-β-D-arabinohexopyranosyl)uracil (S) was obtained in 87% yield. Raney Ni reduction of 8 afforded the corresponding 3′-amino-nucleoside 9. Acetylation of 8 followed by NaBH₄ treatment afforded an 8:1 mixture from which 1-(2,3,4-trideoxy-3-nitro-β-D-threohexopyranosyl)-uracil (14) was obtained in pure crystalline form. After Raney Ni reduction of the mixture, 1-(3-amino-2,3,4-trideoxy-β-d-threo-hexopyranosyl)uracil (16) and its erythro epimer 21 were isolated. 1-(4,6-O-Benzylidene-2,3-dideoxy-3-nitro-β-d-lyxohexopyranosyl)uracil (24) was prepared in 72% yield from 1-(4,6-O-benzylidene-3-deoxy-3-nitro-β-d-galactopyranosyl)uracil (4) by acetylation and subsequent reduction with NaBH₄. De-O-benzylid-enation of 23 afforded 1-(2,3,4-trideoxy-3-nitro-β-d-lyxohexopyranosyl)uracil (25) in 83% yield. Schmidt-Rutz reaction of 25 followed by NaBH₄ reduction afforded a mixture of threo and elythro isomers of 2′,3′,4′-trideoxy-3′-nitro-hexopyranosyluracil, from which pure 16 and 21 were obtained.

     

Solution NMR Conformational Analysis of the Potent Equilibrative Sensitive (ES) Nucleoside Transporter Inhibitor,S 6-(4-Nitrobenzyl)mercaptopurine Riboside (NBMPR)

Abstract

High resolution NMR analysis involving one-dimensional (1-D) ¹H and nuclear Overhauser (NOE) difference spectroscopy was applied to solutions of NBMPR in DMSO-d ₆. Coupling constants were obtained at different temperatures between 285 and 353 K, and used to analyze the rotamer preferences about the C-4′-C-5′ bond. The results revealed a rotamer distribution about the χ tortion angle that favors the high-anti range, a preponderance of the γ⁺ rotamer (at ～64 %) with respect to the γ torsion angle, and a higher population of the south (S) conformer, which was favored by as little as the 4 % to as much as 31 % over the north (N) conformer as calculated by the program PSEUROT 6.2. The high-anti glycosidic torsion orientation appears to be the major conformational difference between the solution structure of NBMPR determined in this study and the structure previously observed in the solid state.     

Ulva rigida (Ulvales,Chlorophyta) tank culture as biofilters for dissolved inorganic nitrogen from fishpond effluents

Ulva rigida was cultivated in 7501 tanks at different densities with direct and continuous inflow (at 2, 4, 8 and 12 volumes d^–1) of the effluents from a commercial marine fishpond (40 metric tonnes, Tm, of Sparus aurata, water exchange rate of 16 m³ Tm^–1) in order to assess the maximum and optimum dissolved inorganic nitrogen (DIN) uptake rate and the annual stability of the Ulva tank biofiltering system. Maximum yields (40 g DW m^–2 d^–1) were obtained at a density of 2.5 g FW 1^–1 and at a DIN inflow rate of 1.7 g DIN m^–2 d^–1. Maximum DIN uptake rates were obtained during summer (2.2 g DIN M^–2 d^–1), and minimum in winter (1.1 g DIN m^–2 d^–1) with a yearly average DIN uptake rate of 1.77 g DIN m^–2 d^–1 At yearly average DIN removal efficiency (2.0 g DIN m^–2 d^–1, if winter period is excluded), 153 m² of Ulva tank surface would be needed to recover 100% of the DIN produced by 1 Tm of fish.Abbreviations DIN= dissolved inorganic nitrogen (NH _inf4 ^sup+ + NO _inf3 ^sup– + NO _inf2 ^sup– ); - FW= fresh weight; - DW= dry weight; - PFD= photon flux density; - V= DIN uptake rate     

Kinetic studies on the reactions of Fusarium galactose oxidase with five different substrates in the presence of dioxygen

 Reactions (25  °C) of galactose oxidase, GOase_ox from Fusarium NRRL 2903 with five different primary-alcohol-containing substrates RCH₂OH:- D-galactose (I) and 2-deoxy-d-galactose (II) (monosaccharides); methyl-β-d-galactopyranoside (III) (glycoside);d-raffinose (IV) (trisaccharide); and dihydroxyacetone (V) have been studied in the presence of O₂. The GOase_ox state has a tyrosyl radical coordinated at a square-pyramidal Cu^II active site, and is a two-equivalent oxidant. Reactant concentrations were [GOase_ox] (0.8–10 μM), RCH₂OH (1.0–6.0 mM), and O₂ (0.14–0.29 mM), with I=0.100 M (NaCl). The reactions, monitored at 450 nm by stopped-flow spectrophotometry, terminated with depletion of the O₂. Each trace was fitted to the competing reactions GOase_ox+RCH₂ OH → GOase_redH₂+RCHO (k ₁), and GOase_redH₂+O₂→ GOase_ox+H₂O₂ (k ₂), with GOase_redH₂ written as the doubly protonated two-electron-reduced Cu^I product. It was necessary to avoid auto-redox interconversion of GOase_ox and GOase_semi . Information obtained at pH 7.5 indicates a 5 : 95 (ox : semi) "native" mix equilibration complete in ∼3 h. At pH >7.5, rate constants 10^–4 k ₁ / M^–1 s^–1 for the reactions of GOase_ox with (I) (1.19), (II) (1.07), (III) (1.29), (IV) (1.81), (V) (2.94) were determined. On decreasing the pH to 5.5, k ₁ values decreased by factors of up to a half, and acid dissociation pK _as in the range 6.6–6.9 were obtained. UV-Vis spectrophotometric studies on GOase_ox gave an independently determined pK _a of 6.7. No corresponding reactions of the Tyr495Phe variant were observed, and there are no similar UV-Vis absorbance changes for this variant. The pK _a is therefore assigned to protonation of Tyr-495 which is a ligand to the Cu. The rate constant k ₂ (1.01×10⁷ M^–1 s^–1) is independent of pH in the range 5.5–9.0 investigated, suggesting that H⁺ (or H-atoms) for the O₂ → H₂O₂ change are provided by the active site of GOase_red . The Cu^I of GOase_red is less extensively complexed, and a coordination number of three is likely. Received: 4 February 1997 / Accepted: 16 May 1997     

Net methane emissions from grazing sheep

Methane emissions from ruminant livestock are responsible for 45 % of New Zealand’s combined CO₂-equivalent greenhouse gas inventory, and arise principally from sheep. Using a flock of 6-month old sheep (20 ha^–1) grazing abundant pasture, we compare micrometeorological measurements of net methane emission rates with measurements from individual sheep based on a sulphur-hexafluoride tracer technique. Individual sheep emission rates were highly variable and averaged 19.5 ± 4.8 (SD) g CH₄ sheep^–1 d^–1, or 39 ± 9.6 mg CH₄ m^–2 d^–1 on an areal basis. Emission rates were poorly correlated with animal live weight or dry matter intake but represented an average dietary energy loss of 3.6%. Methane fluxes from the surface were determined as half hourly averages by a flux-gradient technique using temperature and methane gradients. Soil methane consumption was measured using chambers and found to be negligible (< 0.09 mg CH₄ m^–2 d^–1) in comparison with the animal contribution. Daily net emission rates averaged 46 mg m^–2 d^–1 and exhibited a broad peak in the early afternoon which corresponded with animal activity patterns. On average, net emisssion rates were 40% higher during the day than at night. Stable nocturnal conditions led to a separation of the micrometeorological measurements from the methane source and hence highly variable results. Based on two corroborating techniques, the average net methane emission rate was c. 43 mg CH₄ m^–2 d^–1 or 155 kg CH₄ ha^–1 y^–1.     

In position 7 l- and d-Tic-substituted oxytocin and deamino oxytocin: NMR study and conformational insights

Incorporation of l- or d-Tic into position 7 of oxytocin (OT) and its deamino analogue ([Mpa¹]OT) resulted in four analogues, [l-Tic⁷]OT (1), [d-Tic⁷]OT (2), [Mpa¹,l-Tic⁷]OT (3) and [Mpa¹,d-Tic⁷]OT (4). Their biological properties were described by Fragiadaki et al. (Eur J Med Chem 42:799–806, 2007). Their NMR study (NOESY, TOCSY, ¹H–¹³C HSQC spectra) is presented here. Analogues 1, 3 and 4 showed partial agonistic activity, analogue 2 was pure antagonist, suggesting that a cis conformation between residues 6 and 7 of the molecule does not result in antagonistic activity. However, the reduction in agonistic activity of analogues 1, 3 and 4 in comparison to oxytocin is consistent with the reduction of the trans conformation form. Binding affinity for the human oxytocin receptor with IC₅₀ value of 130, 730, 103, and 380 nM for peptides 1, 2, 3, and 4, respectively, showed lower affinity in the case of d analogues. Deamination slightly increased the affinity. The existence of both cis and trans configurations of the Cys⁶-d-Tic⁷ bond is supported by observation of two sets of cross-peaks for ¹H and ¹³C nuclei for most of the residues of the peptide not only in NOESY and TOCSY but also in ¹H–¹³C HSQC spectra. The MS and HPLC indicate the presence of a single molecule/peptide, and NMR data thus suggest that this second set of peaks is due to the cis conformation.     

The synthesis and use of a 15N and 2H isotopically-labelled derivative of the spin-trap 3, 5-dibromo-4-nitrosobenzenesulphonic acid

Summary

We report the synthesis and use of d₂-¹⁵N isotopically-labelled 3, 5-dibromo-4-nitrosobenzenesulphonic acid (DBNBS-d₂-¹⁵N, as its sodium salt), a spin-trap possessing several advantages over non-labelled DBNBS. The simplification in the electron paramagnetic resonance spectra of radical adducts of DBNBS-d₂-¹⁵N compared with those of DBNBS not only results in increased sensitivity, but also facilitates the assignment and analysis of complex spectra. An example of this simplification is given.     

Dissolved oxygen and nutrient budgets in a phytotreatment pond colonised by <Emphasis Type="Italic">Ulva</Emphasis> spp.

Net daily budgets of dissolved oxygen (O₂), dissolved inorganic carbon (DIC), dissolved inorganic nitrogen (DIN = NH₄⁺+NO₂⁻+NO₃⁻) and soluble reactive phosphorus (SRP) were determined in a pond colonised by Ulva spp. This pond received wastewater from a land-based fish farm and was used as a phytotreatment plant. Three consecutive 24-h cycles of measurements were performed with 8–14 samplings per day. Water samples were collected at the inlet and outlet of the pond and budgets were estimated from differences between inlet and outlet loadings. The first cycle was started when Ulva biomass was 8 kg m⁻², as wet weight. The second cycle was performed after the harvest of ~20% of the macroalgal biomass and the third after the harvest of another ~20% of the remaining biomass. Ulva removal was very fast (<1 h) and samplings for cycles 2 and 3 were started two hours after harvesting, so that the whole experiment lasted ~80 h. When Ulva biomass was at its maximum, the aquatic system was heterotrophic with an O₂ demand of 519 mol d⁻¹ and a net regeneration of DIC (2686 mol d⁻¹), NH₄⁺ (49 mol d⁻¹) and SRP (2.5 mol d⁻¹). The DIC to O₂ ratio was an indicator of persistent anaerobic metabolism. Following the first harvest intervention, this system displayed a prompt response and shifted toward a lower O₂ demand (from −519 to −13 mol d⁻¹), with a lesser regeneration degree of NH₄⁺ (11.4 mol d⁻¹) and DIC (1066 mol d⁻¹). After the second Ulva removal the net budget of SRP became negative (−1.0 mol d⁻¹). By integrating these results over the three days cycle we estimated that in order to operate an efficient nutrient control and maintain macroalgal mats in a healthy status the optimal Ulva biomass should be well below ~4 kg m⁻² as wet weight. Above this threshold, self-limitation would render most of the algal mat unable to exploit light and nutrients. An efficient removal of nitrogen and phosphorus could be attained through the management of macroalgal biomass only with an optimisation of recipient surface to nutrient loading ratio.     

DNA binding,prominent photonuclease activity and antibacterial PDT of cobalt(II) complexes of phenanthroline based photosensitizers

Abstract

The chemistry of Co(II) complexes showing efficient light induced DNA cleavage activity, binding propensity to calf thymus DNA and antibacterial PDT is summarized in this article. Complexes of formulation [Co(mqt)(B)₂]ClO₄ 1–3 where mqt is 4-methylquinoline-2-thiol and B is N,N-donor heterocyclic base, viz. 1,10-phenanthroline (phen 1), dipyrido[3,2-d:2′,3′-f]quinoxaline (dpq 2) and dipyrido[3,2-a:2′,3′-c]phenazine (dppz 3) have been prepared and characterized. The DNA-binding behaviors of these three complexes were explored by absorption spectra, viscosity measurements and thermal denaturation studies. The DNA binding constants for complexes 1, 2 and 3 were determined to be 1.6?×?10³?M^?1, 1.1?×?10⁴?M^?1 and 6.4?×?10⁴?M^?1 respectively. The experimental results suggest that these complexes interact with DNA through groove binding mode. The complexes show significant photocleavage of supercoiled (SC) DNA proceeds via a type-II process forming singlet oxygen as the reactive species. Antimicrobial photodynamic therapy was studied using photodynamic antimicrobial chemotherapy (PACT) assay against E. coli and all complexes exhibited significant reduction in bacterial growth on photoirradiation.     

Mass spectrometric quantification of the mu opioid receptor agonist Tyr-d-Arg-Phe-Lys-NH2 (DALDA) in high-performance liquid chromatography-purified ovine plasma

The mu opioid receptor agonist Tyr-d-Arg-Phe-Lys-Amide (d-Arg²-Lys⁴-Dermorphin_1-4amide=DALDA) was infused continuously for 2 h into sheep. The presence of DALDA in ovine plasma was determined by reversed-phase high-performance liquid chromatography (RP-HPLC) and mass spectrometry (MS) in plasma samples that were obtained at different times during and following that infusion. A stable isotope-incorporated internal standard, deuterated DALDA (d₅-DALDA), was used for the MS quantification of DALDA via the protonated molecule ion, (M+H)⁺, of DALDA and of d₅-DALDA. Time-course data (μg DALDA ml⁻¹ plasma vs. time) were obtained. Tandem MS (MS–MS) provided the product-ion spectrum of the (M+H)⁺ ion of DALDA in one of the samples to confirm the amino acid sequence of DALDA.     

Synthesis and Antimicrobial Evaluation of Novel Pyrazolones and Pyrazolone Nucleosides

The synthesis of a novel series of 4-arylhydrazono-5-methyl-1,2-dihydropyrazol-3-ones 4a–h, and their N ²-alkyl and acyclo, glucopyranosyl, and ribofuranosyl derivatives is described. K₂CO₃ catalyzed alkylation of 4a–h with allyl bromide, propargyl bromide, 4-bromobutyl acetate, 2-acetoxyethoxymethyl bromide, and 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide proceeded selectively at the N ²-position of the pyrazolinone ring. Glycosylation of 4a with 1,2,3,5-tetra-O-acetyl-β-D-ribofuranose under Vorbruggen glycosylation conditions gave the corresponding N ²-4-arylhydrazonopyrazolone ribofuranoside 9a in good yield. Conventional deprotection of the acetyl protected nucleosides furnished the corresponding 4-arylhydrazonopyrazolone nucleosides in good yields. Selected numbers of the newly synthesized compounds were screened for antimicrobial activity. Compounds 4b, 12a, and 14d showed moderate activities against Aspergillus flavus, Penicillium sp., and Escherichia coli.     

Aerobic biodegradation of a mixture of sulfonated azo dyes by a bacterial consortium immobilized in a two‐stage sparged packed‐bed biofilm reactor

The biodegradation of the sulfonated azo dyes, Acid Orange 7 (AO7) and Acid Red 88 (AR88), by a bacterial consortium isolated from water and soil samples obtained from sites receiving discharges from textile industries, was evaluated. For a better removal of azo dyes and their biodegradation byproducts, an aerobically operated two‐stage rectangular packed‐bed biofilm reactor (2S‐RPBR) was constructed. Because the consortium's metabolic activity is affected by oxygen, the effect of the interstitial air flow rate Q_GI on 2S‐RPBR's zonal values of the oxygen mass transfer coefficient k_La was estimated. In the operational conditions probed in the bioreactor, the k_La values varied from 3 to 60 h^?1, which roughly correspond to volumetric oxygen transfer rates, dc_L/dt, ranging from 20 to 375 mg O₂ L^?1h^?1. Complete biodegradation of azo dyes was attained at loading rates B_V,AZ up to 40 mg L^?1d^?1. At higher B_V,AZ values (80 mg L^?1 d^?1), dye decolorization and biodegradation of the intermediaries 4‐amino‐naphthalenesulphonic acid (4‐ANS) and 1‐amino‐2‐naphthol (1‐A2N) was almost complete. However, a diminution in COD and TOC removal efficiencies was observed in correspondence to the 4‐aminobenzenesulfonic acid (4‐ABS) accumulation in the bioreactor. Although the oxygen transport rate improved the azo dye mineralization, the results suggest that the removal efficiency of azo dyes was affected by biofilm detachment at relatively high Q_GI and B_V,AZ values. After 225 days of continuous operation of the 2S‐RFBR, eight bacterial strains were isolated from the biofilm attached to the porous support. The identified genera were: Arthrobacter, Variovorax, Agrococcus, Sphingomonas, Sphingopyxis, Methylobacterium, Mesorhizobium, and Microbacterium.