Synthesis of 2-Deoxy-β-D-ribose 1-Phosphate,NMR Comparison and Its Enzymatic Activity for Structural Elucidation of Synthetic α-Isomer

Abstract

2-Deoxy-β-D-ribose 1-phosphate (1) was synthesized in a stereoselective manner and isolated with no detectable contamination by its α-isomer (4). Explicit configuration of 4 was first determined by NMR comparison with 1 judging from NOE results and their coupling constants. Natural purine nucleoside phosphorylase (PNPase) did not recognize 1 and gave no products such as α- or β-deoxynucleosides.     

Nucleosides and Related Substances

A new procedure which involves 1-trichloroacetyl sugars as the starting material has been developed for the synthesis of purine nucleosides. 7-β-d-Glucopyranosyl-, 7-β-d-xylopyranosyl-, 7-β-d-ribopyranosyl-theophylline, 9-(tetra-O-acetyl-β-d-glucopyranosyl)-2,6,8-trichloropurine and 9-β-d-glucopyranosyl adenine were prepared in good yields by the reaction in fusion of purine bases with 1-trichloroacetyl sugars, using zinc chloride, p-toluenesulfonic acid, or ethyl polyphosphate as catalyst. 9-d-Ribofuranosyl adenine was also prepared by the same procedures, although the anomeric configuration of the compound is not yet definite. The effect of catalysts on the yields of purine nucleosides is discussed.     

L-Tyrosine production by Polyauxotrophic Mutants of Corynebacterium glutamicum

Polyauxotrophic mutants of Corynebacterium glutamicum which have additional requirements to L-phenylalanine were derived from L-tyrosine producing strains of phenylalanine auxotrophs, C. glutamicum KY 9189 and C. glutamicum KY 10233, and screened for L-tyrosine production. The increase of L-tyrosine production was noted in many auxotrophic mutants derived from both strains. Especially some double auxotrophs which require phenylalanine and purine, phenylalanine and histidine, or phenylalanine and cysteine produced significantly higher amounts of L-tyrosine compared to the parents, A phenylalanine and purine double auxotrophic strain LM–96 produced L-tyrosine at a concentration of 15.1 mg per ml in the medium containing 20% sucrose. L-Tyrosine production by the strain decreased at high concentrations of L-phenylalanine.     

Synthesis and Biological Evaluation of Some 5-Nitro- and 5-Amino Derivatives of 2′-Deoxycytidine, 2′,3′-Dideoxyuridine,and 2′,3′-Dideoxycytidine

Abstract

In this article, we describe the synthesis of 5-nitro-1-(2-deoxy-α-D-erythro-pentofuranosyl)cytosine (4α), 5-nitro-1-(2-deoxy-β-D-erythro-pentofuranosyl)cytosine (4β), 5-amino-1-(2-deoxy-α-D-erythro-pentofuranosyl)cytosine (5α), 5-nitro-1- (2-deoxy-β-D-erythro-pentofuranosyl)cytosine (5β), 5-nitro-1-(2,3-dideoxy-β- D-ribofuranosyl)uracil (6β), 5-amino-1-(2,3-dideoxy-α,β-D-ribofuranosyl)uracil (7), 5-nitro-1-(2,3-dideoxy-α,β-D-ribofuranosyl)cytosine (8) and 5-amino-1-(2,3-dideoxy-β-D-ribofuranosyl)cytosine (9β). The prepared compounds were tested for their activity against HIV and HBV viruses, but they did not show significant activity.     

Syntheses of Several β-l-Rhamnopyranosides

To investigate the substrate specificity of β-l-rhamnosidase, the following β-l-rhamnopyranosides were synthesized: 1-(β-l-rhamnopyranosyl)-dl-glycerol (1), methyl β-l-rhamnopyranoside (2), methyl 2-O-(β-l-rhamnopyranosyl)-β-d-glucopyranoside (3) and methyl 2-O-β(β-l-rhamnopyranosyl)-α-l-arabinopyranoside (4). The synthesis of 3 was performed using l-quinovose with neighboring group participation, which lead stereoselectively to the β-l-quinovoside. The 2-OH of the l-quinovo-unit was selectively deblocked, oxidized to the keto group, and then stereoselectively reduced, whereby 3 was produced.     

DNA binding,photocleavage, antimicrobial and cytotoxic properties of Ru(II) polypyridyl complexes containing BOPIP ligand, (BOPIP = {2-(4-(benzyloxy) phenyl)-1H-imidazo [4,5-f] [1,2]phenanthroline})

A novel ligand BOPIP (BOPIP?=?{2-(4-(benzyloxy)phenyl)-1H-imidazo[4,5-f][1,10]phenanthroline}) and its mononuclear Ru(II) polypyridyl complexes [Ru(phen)₂ BOPIP]²⁺(1) (phen?=?1,10-Phenanthrolene), [Ru(bpy)₂ BOPIP]²⁺(2) (bpy?=?2,2′ bipyridyl), [Ru(dmb)₂ BOPIP]²⁺(3) (dmb?=?4, 4′ -dimethyl 2, 2′ -bipyridine), [Ru(Hdpa)₂ BOPIP]²⁺(4) (Hdpa?=?2,2′dipyridylamine) have been synthesized successfully and characterized by elemental analysis, UV-vis, IR, ¹H, ¹³ Gill, M. R.; Garcia-Lara, J.; Foster, S. J.; Smythe, C.; Battaglia, G.; Thomas, J. A. A Ruthenium(II) polypyridyl Complex for Direct Imaging of DNA Structure in Living Cells. Nat. Chem. 2009, 1, 662–667.[Crossref], [PubMed], [Web of Science ®] , [Google Scholar]C-NMR, and ESI-MS Spectroscopy. The interaction of these complexes with CT-DNA was studied using absorption, emission techniques, viscosity measurements and molecular docking studies. The docking study also supports the binding ability of complexes obtained through the absorption and emission techniques. These studies reveal that the Four Ru(II) polypyridyl complexes bind to DNA predominantly by intercalation. The Antimicrobial activity and cytotoxicity of these complexes are also reported.     

Carnivore Trackways from the Miocene Site of Salinas de Añana (Alava,Spain)

A fossil tracksite of Lower Miocene age discovered near Salinas de Añana, Alava (Spain), has rendered an exceptionally well-preserved assemblage of vertebrate ichnites. The site shows a high proportion of carnivore tracks (3 out of 5 mammal ichnospecies) and a high number of individual trackways (15), some including over 50 consecutive footprints. The carnivore ichnites are classified as Felipeda lynxi Panin & Avram, 1962 Panin, M. and Avram, B. 1962. Noi urme de vertebrate in Miocenul Subcarpatilor Rominesti. Studii Cercet Geologia, 13: 521–525.  [Google Scholar], Felipeda parvula ichnosp. nov. and Canipeda longigriffa Panin & Avram, 1962 Panin, M. and Avram, B. 1962. Noi urme de vertebrate in Miocenul Subcarpatilor Rominesti. Studii Cercet Geologia, 13: 521–525.  [Google Scholar], and they are attributed to a felid, an undetermined small aeluroid, and a herpestid, respectively. The long trackways allow determination of gaits, which include lateral sequence walks and diagonal sequence trots, and of speed, which ranges from 0.4 to 1.4 m/s. Froude numbers range between 0.1 and 0.8, agreeing with gait interpretations and speed calculations. The felid trackways provide the first known evidence of group traveling in fossil cats. The herpestid footprints show modern-grade adaptations for terrestrial locomotion and digging.     

Optimization of the 503 antigen induction strategy of Leishmania infantum chagasi expressed in Escherichia coli M15

Abstract

Leishmaniosis is a complex of diseases that can be fatal, if not given proper attention. Despite its relevance in the public health system, there is no vaccine capable of preventing the disease in humans so far and its treatment is expensive and aggressive to human health. The present study aims to optimize the induction parameters of the 503 Leishmania i. chagasi antigen expressed in recombinant Escherichia coli M15. The induction at different cell densities was evaluated in order to analyze the influence of the induction time on the yield of the protein of interest. In this segment, lactose and isopropyl-β-d-thiogalactopyranoside (IPTG) were used as inducer molecules, using various concentrations: 0.1?g/L, 1.0?g/L, and 10?g/L for lactose and 20?μM, 100?μM, 500?μM, and 1000?μM for IPTG. The results presented that the concentration of IPTG that obtained the higher antigen levels was that of 100?μM (0.087?g/L), a 10-fold lower concentration than was being previously used in this type of system and for lactose, it was 1?g/L (0.016?g/L). Thus, the induction with 100?μM allowed obtaining the antigen with a concentration 5.6 times higher than the lactose induction maximum concentration.     

A New Synthesis of 9-(2-Deoxy-2-fluoro-β-D-arabinofuranosyl)guanine (araF-G)

Abstract

Interesting and very promising antisense properties of 2′-deoxy-2′-fluoroarabinonucleic acids ((a) Wilds, C.J.; Damha, M.J. 2′-Deoxy-2′-fluoroarabinonucleosides and oligonucleotides (2′F-ANA): synthesis and physicochemical studies. Nucl. Acids Res. 2000, 28, 3625–3635; (b) Viazovkina, E.; Mangos, M.; Elzagheid, M.I.; Damha, M.J. Current Protocols in Nucleic Acid Chemistry 2002, 4.15.1–4.15.21) (2′F-ANA) has encouraged our research group to optimize the synthetic procedures for 2′-deoxy-2′-fluoro-β-D-arabinonucleosides (araF-N). The synthesis of araF-U, araF-T, araF-A and araF-C is straightforward, (Tann, C.H.; Brodfuehrer, P.R.; Brundidge, S.P.; Sapino, C., Jr. Howell H.G. Fluorocarbohydrates in synthesis. An efficient synthesis of 1-(2-deoxy-2-fluoro-β-D-arabinofuranosyl)-5-iodouracil (β-FIAU) and 1-(2-deoxy-2-fluoro-β-D-arabinofuranosyl)thymine (β-FMAU). J. Org. Chem. 1985, 50, 3644–3647; Howell, H.G.; Brodfuehrer, P.R.; Brundidge, S.P.; Benigni, D.A.; Sapino, C., Jr. Antiviral nucleosides. A stereospecific, total synthesis of 2′-fluoro-2′-deoxy-β-D-arabinofuranosyl nucleosides. J. Org. Chem. 1988, 53, 85–88; Maruyama, T.; Takamatsu, S.; Kozai, S.; Satoh, Y.; Izana, K. Synthesis of 9-(2-deoxy-2-fluoro-β-D-arabinofuranosyl)adenine bearing a selectively removable protecting group. Chem. Pharm. Bull. 1999, 47, 966–970) however, the synthesis of the guanine analogue is more complicated and affords poor to moderate yields of araF-G (4) ((a) Elzagheid, M.I.; Viazovkina, E.; Masad, M.J. Synthesis of protected 2′-deoxy-2′-fluoro-β-D-arabinonucleosides. Synthesis of 2′-fluoroarabino nucleoside phosphoramidites and their use in the synthesis of 2′F-ANA. Current Protocols in Nucleic Acid Chemistry 2002, 1.7.1–1.7.19; (b) Tennila, T.; Azhayeva, E.; Vepsalainen, J.; Laatikainen, R.; Azhayev, A.; Mikhailopulo, I. Oligonucleotides containing 9-(2-deoxy-2-fluoro-β-D-arabinofuranosyl)-adenine and -guanine: synthesis, hybridization and antisense properties. Nucleosides, Nucleotides and Nucl. Acids 2000, 19, 1861–1884). Here we describe an efficient synthesis of araF-G (4) that involves coupling of 2-deoxy-2-fluoro-3,5-di-O-benzoyl-α-D- arabinofuranosyl bromide (1) with 2-chlorohypoxanthine (2) to afford 2-chloro-β-araF-I (3) in 52% yield. Nucleoside (3) was transformed into araF-G (4) by treatment with methanolic ammonia (150°C, 6 h) in 67% yield.     

Oligonucleotides Containing Disaccharide Nucleosides: Synthesis,Physicochemical, and Substrate Properties

Abstract

The efficient synthesis of oligonucleotides containing 2′-O-β-D-ribofuranosyl (and β-D-ribopyranosyl)nucleosides, 2′-O-α-D-arabinofuranosyl (and α-L-arabinofuranosyl)nucleosides, 2′-O-β-D-erythrofuranosylnucleosides, and 2′-O-(5′-amino-5-deoxy-β-D-ribofuranosyl)nucleosides have been developed.     

Chemo-enzymatic Synthesis of 3-Deoxy-β-D-ribofuranosyl Purines and Study of Their Biological Properties

Abstract

9-(3-Deoxy-β-d-erythro-pentofuranosyl)-2,6-diaminopurine (2) was synthesized by an enzymatic transglycosylation of 2,6-diaminopurine using 3′-deoxycytidine (1) as a donor of the sugar moiety. Nucleoside 2 was transformed to 3′-deoxy guanosine (3), 9-(3-deoxy-β-d-erythro-pentofuranosyl)-2-amino-6-oxopurine (3′-deoxyisoguanosine; 4), and 9-(3-deoxy-β-d-erythro-pentofuranosyl)-2-fluoroadenine (5). Compounds 2–5 were evaluated for their anti-HIV activity.     

MOLECULAR MODELLING OF 2′-OH MEDIATED HYDROGEN BONDING IN RIBONUCLEOS(T)IDES BY NMR CONSTRAINED AM1 AND MMX CALCULATIONS

The intra- and intermolecular hydrogen bonding (ΔGº_298K ≈ 2, kcal mol^?1) of 2′-OH in nucleos(t)ides has been reported by the temperature- and concentration-dependent NMR study in conjunction with dihedral dependence of the NMR derived both endo (³ J _H,H)- and exocyclic (³ JH,OH) coupling constants, nOe contacts and lineshape analyses of hydroxyl protons for EtpA (1), 3′-dA (2), rA (3), 2′-dA (4) [Fig. 1] in DMSO-d ₆ at 500 MHz.

MOLECULAR MODELLING OF 2′-OH MEDIATED HYDROGEN BONDING IN RIBONUCLEOS(T)IDES BY NMR CONSTRAINED AM1 AND MMX CALCULATIONS

All authors

Parag Acharya, Irina Velikian, Sandipta Acharya & Jyoti Chattopadhyaya

https://doi.org/10.1081/NCN-100002521

Published online:

07 February 2007

Figure 1. The schematic representation of the bias of the dymanic two-state pseudorotational equilibrium between the North-type (N, C2′-exo -C3′-endo) and the South-type (S, C3′-exo-C2′-endo) [3a] Thibaudeau, C. and Chattopadhyaya, J. 1999. Stereoelectronic Effects in Nucleosides and Nucleotides and their Structural Implications Sweden: (ISBN 91-506-1351-0), Department of Bioorganic Chemistry, Uppsala University Press (fax: +4618554495). For review see: and references therein [Google Scholar] pseudorotamers of the sugar moeity for EtpA (1), 3′-dA (2), rA (3), 2′-dA (4) and torsion (Φ) around C2′/3′-O bond viz. Φ₁ = Φ_{H2′?C2′?O?H} and Φ₂ = Φ_{H3′?C3′?O?H} except in 1 where the torsion across C3′-O3′ bond is actually ?^? [C4′-C3′-O3′-P].     

14.

Molecular identity,morphology and taxonomy of the Rhynchospio glutaea complex with a key to Rhynchospio species (Annelida,Spionidae)     

Vasily I. Radashevsky  Tatyana V. Neretina  Victoria V. Pankova  Alexander B. Tzetlin  Jin-Woo Choi 《分类学与生物多样性》2013,11(4):424-433

Rhynchospio glutaea (Ehlers, 1897 Ehlers, E. 1897. Polychaeten. Ergebnisse der Hamburger Magalhaensischen Sammelreise 1892/93. L. Friederichsen & Co., Hamburg, Germany. [Google Scholar]), Rhynchospio arenincola Hartman, 1936 Hartman, O. 1936. New species of Spionidae (Annelida Polychaeta) from the coast of California. University of California Publications in Zoology 41, 45–52. [Google Scholar] and Rhynchospio arenincola asiatica Chlebovitsch, 1959 Chlebovitsch, V.V. 1959. Species of Polychaeta worms from the Kurile Islands, which are new or recorded for the first time in the USSR fauna. Zoologichesky Zhurnal 38, 167–181. [Google Scholar], originally described from Strait of Magellan, California, and South Kurile Islands respectively, appear similar to each other in adult morphology. These species and subspecies have been considered by some authors as subjective synonyms and are here referred to as members of the R. glutaea complex. Sequence data of four gene fragments (2465 bp in total) of the mitochondrial 16S rDNA, nuclear 18S and 28S rDNA, and Histone 3 have shown that R. glutaea complex individuals from the South West Atlantic (Argentina), North East Pacific (British Columbia and Oregon) and North West Pacific (South Korea) were genetically distant and not conspecific. These data also indicate that R. arenincola from North America and R. aff. asiatica from South Korea are more closely related to each other, and both are closer to R. glutaea from South America than to R. nhatrangi from Vietnam: nhatrangi (glutaea (arenincola-aff. asiatica)). Adults of the R. glutaea complex are hermaphrodites and the arrangement of gametes is suggested to be a crucial reproductive character for distinguishing these species. Based on this character, two species of the complex are apparent in the North West Pacific: R. asiatica Chlebovitsch, 1959 Chlebovitsch, V.V. 1959. Species of Polychaeta worms from the Kurile Islands, which are new or recorded for the first time in the USSR fauna. Zoologichesky Zhurnal 38, 167–181. [Google Scholar] stat. nov. from the Kurile Islands (not analysed here) and an unnamed species from the mainland coast of Asia (here referred to as R. aff. asiatica). Adult morphology of R. asiatica stat. nov. is briefly described and illustrated. The lectotype and the type locality of the species on Iturup Is. are established for the first time. An identification key is provided to the 10 currently recognized Rhynchospio species.     

15.

NOVEL SYNTHESIS OF seco TYPE OF ACYCLO C-NUCLEOSIDES OF 1,2,4-TRIAZOLE AND 1,2,4-TRIAZOLO[3,4-b][1,3,4]THIADIAZINE     

《Nucleosides, nucleotides & nucleic acids》2013,32(1-2):103-116

The seco C-nucleosides 3-(1,2,3,4,5-penta-O-acetyl-D-gluco- and D- galacto-pentitol-1-yl)-1H-1,2,4-triazoles (8 and 9) were obtained in a one pot by deamination and dethiolation of 4-amino-3-(D-gluco- and D-galacto-pentitol-1-yl)-5-mercapto-1,2,4-triazoles (1 and 2), respectively, using sodium nitrite in orthophosphoric acid and subsequent acetylation. Condensation of 1, 2, and 4-amino-3-(D-glycero-D-gulo-hexitol-1-yl)-5-mercapto-1,2,4-triazole (12) with phenacylbromide (11) afforded the corresponding 3-(D-gluco-, D-galacto-pentitol-1-yl) and 3-(D-glycero-D-gulo-hexitol-1-yl)-6-phenyl-7H-1,2,4- triazolo[3,4-b][1,3,4] thiadiazines (15, 16, and 17). Acetylation of 15–17 gave the penta- and hexa-O-acetyl derivatives 18–20, respectively. The structures were confirmed by using ¹H, ¹³C, and 2D NMR spectra, DQFCOSY, HMQC, and HMBC experiments. The favored conformational structures were deduced from the vicinal coupling constants of the protons.     

16.

Neotropical Osmylidae larvae (Insecta,Neuroptera): description of habitats and morphology     

Caleb Califre Martins  Adrian Ardila-Camacho  Gregory W. Courtney 《水生昆虫》2018,39(2-3):181-207

ABSTRACT

The larval stages of the genus Kempynus Navás, 1912 Navás, L. (1912), ‘Insectos neurópteros nuevos o poco conocidos’, Memorias de la Real Academia de Ciencias y Artes de Barcelona, 10, 135–202. [Google Scholar] (probably K. falcatus Navás, 1912 Navás, L. (1912), ‘Insectos neurópteros nuevos o poco conocidos’, Memorias de la Real Academia de Ciencias y Artes de Barcelona, 10, 135–202. [Google Scholar] based on the presence of synchronic and sympatric adults) are described for the Neotropical Region for the first time, and the larval stages of Isostenosmylus pulverulentus (Gerstaecker, 1893 Gerstaecker, C.E.A. (1893), ‘Ueber neue und weniger gekannte Neuropteren aus der familie Megaloptera Burm’, Mitt[h]eilungen aus dem Naturwissenschaftlichen Verein für Neu-Vorpommern und Rugen, 25, 93–173. [Google Scholar]) are redescribed. The external morphology of third-instar larvae of both species and their habitats are described and compared. Kempynus sp. is a water-dependent species and can be considered semi-aquatic, whereas I. pulverulentus larvae are terrestrial and live in undergrowth vegetation. The first key to identification of Neotropical Osmylidae larvae is provided, based on third-instar larvae of both species.     

17.

An Improved Synthesis of DL-Canavanine     

Yasuhiro Yamada  Hiroshi Noda  Hirosuke Okada 《Bioscience, biotechnology, and biochemistry》2013,77(9):2201-2203

Excellent l-proline producers were screened for among sulfaguanidine resistant mutants derived from three typical l-glutamic acid-producing bacteria: Brevibacterium flavum, B. lactofermentum, and C. glutamicum.

The best strain, No. 199, is a sulfaguanidine resistant mutant derived from an isoleucine auxotroph of B. flavum 2247 by nitrosoguanidine. Strain No. 199 produced 35 mg/ml of l-proline after 72 hr of cultivation with 10% glucose as a carbon source. The strain also accumulated purine bases such as adenine, guanine, and hypoxanthine, i.e., degradation products of purine nucleotides. In the mutant, 1.6 ~ 2.0 fold more intracellular ATP was found than that in the parent strain; it is a substrate of glutamate kinase relating to l-proline biosynthesis.

On the contrary, the levels of intracellular glutamic acid, a substrate of glutamate kinase, were similar among these strains.

It was confirmed that the increment of internal ATP, which was important in the l-proline production mechanism, was very effective in the improvement of l-proline producers.     

18.

Biochemical characteristics of maltose phosphorylase MalE from Bacillus sp. AHU2001 and chemoenzymatic synthesis of oligosaccharides by the enzyme     

Yu Gao  Wataru Saburi  Yodai Taguchi 《Bioscience, biotechnology, and biochemistry》2013,77(11):2097-2109

ABSTRACT

Maltose phosphorylase (MP), a glycoside hydrolase family 65 enzyme, reversibly phosphorolyzes maltose. In this study, we characterized Bacillus sp. AHU2001 MP (MalE) that was produced in Escherichia coli. The enzyme exhibited phosphorolytic activity to maltose, but not to other α-linked glucobioses and maltotriose. The optimum pH and temperature of MalE for maltose-phosphorolysis were 8.1 and 45°C, respectively. MalE was stable at a pH range of 4.5–10.4 and at ≤40°C. The phosphorolysis of maltose by MalE obeyed the sequential Bi–Bi mechanism. In reverse phosphorolysis, MalE utilized d-glucose, 1,5-anhydro-d-glucitol, methyl α-d-glucoside, 2-deoxy-d-glucose, d-mannose, d-glucosamine, N-acetyl-d-glucosamine, kojibiose, 3-deoxy-d-glucose, d-allose, 6-deoxy-d-glucose, d-xylose, d-lyxose, l-fucose, and l-sorbose as acceptors. The k_cat(app)/K_m(app) value for d-glucosamine and 6-deoxy-d-glucose was comparable to that for d-glucose, and that for other acceptors was 0.23–12% of that for d-glucose. MalE synthesized α-(1→3)-glucosides through reverse phosphorolysis with 2-deoxy-d-glucose and l-sorbose, and synthesized α-(1→4)-glucosides in the reaction with other tested acceptors.     

19.

A Facile Synthetic Method for 3′-α-Fluoro- 2′,3′-dideoxyadenosine     

《Nucleosides, nucleotides & nucleic acids》2013,32(5-8):711-713

Abstract

A facile method for the synthesis of 3′-α-fluoro-2′,3′-dideoxyadenosine (5) has been developed using a novel rearrangement of 3′-β-bromine to the 2′-β position during 3′-α fluorination.     

20.

Properties of Nucleoside Phosphorylase from Enterobacter aerogenes     

Takashi Utagawa  Hirokazu Morisawa  Shigeru Yamanaka  Akihiro Yamazaki  Fumihiro Yoshinaga  Yoshio Hirose 《Bioscience, biotechnology, and biochemistry》2013,77(11):3239-3246

The properties of uridine Phosphorylase (UPase) and purine nucleoside Phosphorylase (PNPase) at high temperature were investigated. Both enzymes were found to be distributed in a wide range of bacteria and were partially purified from Enterobacter aerogenes AJ 11125 by heat treatment, ammonium sulfate fractionation and column chromatographies onDEAE-cellulose and Sephadex G-150. The UPase was purified 109-fold, and it showed an optimum pH of 8.5 and optimum temperature of 65°C, and activity toward uridine, 2′-deoxyuridine, thymidine and uracil arabinoside but not cytidine. The Km values of UPase for uridine were 0.7 mm at 40°C and 1.8 mm at 60°C. The PNPase was purified 83-fold, and it showed an optimum pH of 6.8 and optimum temperature of 60°C, and significant activity toward purine arabinosides as well as purine ribosides. The Km values of PNPase for inosine were 0.8 mm at 40°C and 2.2 mm at 60°C.     

Molecular identity,morphology and taxonomy of the Rhynchospio glutaea complex with a key to Rhynchospio species (Annelida,Spionidae)

Rhynchospio glutaea (Ehlers, 1897 Ehlers, E. 1897. Polychaeten. Ergebnisse der Hamburger Magalhaensischen Sammelreise 1892/93. L. Friederichsen & Co., Hamburg, Germany. [Google Scholar]), Rhynchospio arenincola Hartman, 1936 Hartman, O. 1936. New species of Spionidae (Annelida Polychaeta) from the coast of California. University of California Publications in Zoology 41, 45–52. [Google Scholar] and Rhynchospio arenincola asiatica Chlebovitsch, 1959 Chlebovitsch, V.V. 1959. Species of Polychaeta worms from the Kurile Islands, which are new or recorded for the first time in the USSR fauna. Zoologichesky Zhurnal 38, 167–181. [Google Scholar], originally described from Strait of Magellan, California, and South Kurile Islands respectively, appear similar to each other in adult morphology. These species and subspecies have been considered by some authors as subjective synonyms and are here referred to as members of the R. glutaea complex. Sequence data of four gene fragments (2465 bp in total) of the mitochondrial 16S rDNA, nuclear 18S and 28S rDNA, and Histone 3 have shown that R. glutaea complex individuals from the South West Atlantic (Argentina), North East Pacific (British Columbia and Oregon) and North West Pacific (South Korea) were genetically distant and not conspecific. These data also indicate that R. arenincola from North America and R. aff. asiatica from South Korea are more closely related to each other, and both are closer to R. glutaea from South America than to R. nhatrangi from Vietnam: nhatrangi (glutaea (arenincola-aff. asiatica)). Adults of the R. glutaea complex are hermaphrodites and the arrangement of gametes is suggested to be a crucial reproductive character for distinguishing these species. Based on this character, two species of the complex are apparent in the North West Pacific: R. asiatica Chlebovitsch, 1959 Chlebovitsch, V.V. 1959. Species of Polychaeta worms from the Kurile Islands, which are new or recorded for the first time in the USSR fauna. Zoologichesky Zhurnal 38, 167–181. [Google Scholar] stat. nov. from the Kurile Islands (not analysed here) and an unnamed species from the mainland coast of Asia (here referred to as R. aff. asiatica). Adult morphology of R. asiatica stat. nov. is briefly described and illustrated. The lectotype and the type locality of the species on Iturup Is. are established for the first time. An identification key is provided to the 10 currently recognized Rhynchospio species.     

NOVEL SYNTHESIS OF seco TYPE OF ACYCLO C-NUCLEOSIDES OF 1,2,4-TRIAZOLE AND 1,2,4-TRIAZOLO[3,4-b][1,3,4]THIADIAZINE

The seco C-nucleosides 3-(1,2,3,4,5-penta-O-acetyl-D-gluco- and D- galacto-pentitol-1-yl)-1H-1,2,4-triazoles (8 and 9) were obtained in a one pot by deamination and dethiolation of 4-amino-3-(D-gluco- and D-galacto-pentitol-1-yl)-5-mercapto-1,2,4-triazoles (1 and 2), respectively, using sodium nitrite in orthophosphoric acid and subsequent acetylation. Condensation of 1, 2, and 4-amino-3-(D-glycero-D-gulo-hexitol-1-yl)-5-mercapto-1,2,4-triazole (12) with phenacylbromide (11) afforded the corresponding 3-(D-gluco-, D-galacto-pentitol-1-yl) and 3-(D-glycero-D-gulo-hexitol-1-yl)-6-phenyl-7H-1,2,4- triazolo[3,4-b][1,3,4] thiadiazines (15, 16, and 17). Acetylation of 15–17 gave the penta- and hexa-O-acetyl derivatives 18–20, respectively. The structures were confirmed by using ¹H, ¹³C, and 2D NMR spectra, DQFCOSY, HMQC, and HMBC experiments. The favored conformational structures were deduced from the vicinal coupling constants of the protons.     

Neotropical Osmylidae larvae (Insecta,Neuroptera): description of habitats and morphology

ABSTRACT

The larval stages of the genus Kempynus Navás, 1912 Navás, L. (1912), ‘Insectos neurópteros nuevos o poco conocidos’, Memorias de la Real Academia de Ciencias y Artes de Barcelona, 10, 135–202. [Google Scholar] (probably K. falcatus Navás, 1912 Navás, L. (1912), ‘Insectos neurópteros nuevos o poco conocidos’, Memorias de la Real Academia de Ciencias y Artes de Barcelona, 10, 135–202. [Google Scholar] based on the presence of synchronic and sympatric adults) are described for the Neotropical Region for the first time, and the larval stages of Isostenosmylus pulverulentus (Gerstaecker, 1893 Gerstaecker, C.E.A. (1893), ‘Ueber neue und weniger gekannte Neuropteren aus der familie Megaloptera Burm’, Mitt[h]eilungen aus dem Naturwissenschaftlichen Verein für Neu-Vorpommern und Rugen, 25, 93–173. [Google Scholar]) are redescribed. The external morphology of third-instar larvae of both species and their habitats are described and compared. Kempynus sp. is a water-dependent species and can be considered semi-aquatic, whereas I. pulverulentus larvae are terrestrial and live in undergrowth vegetation. The first key to identification of Neotropical Osmylidae larvae is provided, based on third-instar larvae of both species.     

An Improved Synthesis of DL-Canavanine

Excellent l-proline producers were screened for among sulfaguanidine resistant mutants derived from three typical l-glutamic acid-producing bacteria: Brevibacterium flavum, B. lactofermentum, and C. glutamicum.

The best strain, No. 199, is a sulfaguanidine resistant mutant derived from an isoleucine auxotroph of B. flavum 2247 by nitrosoguanidine. Strain No. 199 produced 35 mg/ml of l-proline after 72 hr of cultivation with 10% glucose as a carbon source. The strain also accumulated purine bases such as adenine, guanine, and hypoxanthine, i.e., degradation products of purine nucleotides. In the mutant, 1.6 ~ 2.0 fold more intracellular ATP was found than that in the parent strain; it is a substrate of glutamate kinase relating to l-proline biosynthesis.

On the contrary, the levels of intracellular glutamic acid, a substrate of glutamate kinase, were similar among these strains.

It was confirmed that the increment of internal ATP, which was important in the l-proline production mechanism, was very effective in the improvement of l-proline producers.     

Biochemical characteristics of maltose phosphorylase MalE from Bacillus sp. AHU2001 and chemoenzymatic synthesis of oligosaccharides by the enzyme

ABSTRACT

Maltose phosphorylase (MP), a glycoside hydrolase family 65 enzyme, reversibly phosphorolyzes maltose. In this study, we characterized Bacillus sp. AHU2001 MP (MalE) that was produced in Escherichia coli. The enzyme exhibited phosphorolytic activity to maltose, but not to other α-linked glucobioses and maltotriose. The optimum pH and temperature of MalE for maltose-phosphorolysis were 8.1 and 45°C, respectively. MalE was stable at a pH range of 4.5–10.4 and at ≤40°C. The phosphorolysis of maltose by MalE obeyed the sequential Bi–Bi mechanism. In reverse phosphorolysis, MalE utilized d-glucose, 1,5-anhydro-d-glucitol, methyl α-d-glucoside, 2-deoxy-d-glucose, d-mannose, d-glucosamine, N-acetyl-d-glucosamine, kojibiose, 3-deoxy-d-glucose, d-allose, 6-deoxy-d-glucose, d-xylose, d-lyxose, l-fucose, and l-sorbose as acceptors. The k_cat(app)/K_m(app) value for d-glucosamine and 6-deoxy-d-glucose was comparable to that for d-glucose, and that for other acceptors was 0.23–12% of that for d-glucose. MalE synthesized α-(1→3)-glucosides through reverse phosphorolysis with 2-deoxy-d-glucose and l-sorbose, and synthesized α-(1→4)-glucosides in the reaction with other tested acceptors.     

A Facile Synthetic Method for 3′-α-Fluoro- 2′,3′-dideoxyadenosine

Abstract

A facile method for the synthesis of 3′-α-fluoro-2′,3′-dideoxyadenosine (5) has been developed using a novel rearrangement of 3′-β-bromine to the 2′-β position during 3′-α fluorination.     

Properties of Nucleoside Phosphorylase from Enterobacter aerogenes

The properties of uridine Phosphorylase (UPase) and purine nucleoside Phosphorylase (PNPase) at high temperature were investigated. Both enzymes were found to be distributed in a wide range of bacteria and were partially purified from Enterobacter aerogenes AJ 11125 by heat treatment, ammonium sulfate fractionation and column chromatographies onDEAE-cellulose and Sephadex G-150. The UPase was purified 109-fold, and it showed an optimum pH of 8.5 and optimum temperature of 65°C, and activity toward uridine, 2′-deoxyuridine, thymidine and uracil arabinoside but not cytidine. The Km values of UPase for uridine were 0.7 mm at 40°C and 1.8 mm at 60°C. The PNPase was purified 83-fold, and it showed an optimum pH of 6.8 and optimum temperature of 60°C, and significant activity toward purine arabinosides as well as purine ribosides. The Km values of PNPase for inosine were 0.8 mm at 40°C and 2.2 mm at 60°C.