Biosynthesis of riboflavin. Single turnover kinetic analysis of 6,7-dimethyl-8-ribityllumazine synthase

6,7-dimethyl-8-ribityllumazine synthase (lumazine synthase) catalyzes the condensation of 5-amino-6-ribitylamino-2,4-(1H,3H)-pyrimidinedione with 3,4-dihydroxy-2-butanone 4-phosphate, affording the riboflavin precursor, 6,7-dimethyl-8-ribityllumazine. Single turnover experiments monitored by multiwavelength photometry were performed with the recombinant lumazine synthase of Bacillus subtilis. Mixing of the enzyme with the pyrimidine type substrate is conducive to a hypsochromic shift as well as a decrease in absorbance of the heterocyclic substrate; the rate constant for that reaction is 0.02 s(-1) microM(-1). Rapid mixing of the complex between enzyme and pyrimidine type substrate with the second substrate, 3,4-dihydroxy-2-butanone 4-phosphate, is followed by the appearance of an early optical transient characterized by an absorption maxima at 330 nm of low intensity which was tentatively assigned as a Schiff base intermediate. The subsequent elimination of phosphate affords a transient with intense absorption maxima at 455 and 282 nm, suggesting an intermediate with an extended system of conjugated double bonds. The subsequent formation of the enzyme product, 6,7-dimethyl-8-ribityllumazine, is the rate-determining step.     

Effects of spiroconjugation on the calculated singlet-triplet energy gap in 2,2-dialkoxycyclopentane-1,3-diyls and on the experimental electronic absorption spectra of singlet 1,3-diphenyl derivatives. Assignment of the lowest-energy electronic transition of singlet cyclopentane-1,3-diyls

The effect of a 2,2-ethylene-ketal functionality on the singlet-triplet energy gap (Delta E(ST)) and on the first electronic transition in singlet cyclopentane-1,3-diyls (1) has been investigated. UDFT calculations predict a significant increase in the preference for a singlet ground state in the diradical with the cyclic ketal at C2 (1g; Delta E(ST) = -6.6 kcal/mol in C(2) symmetry and -7.6 kcal/mol in C(2v) symmetry), compared to the 2,2-dihydroxy- and 2,2-dimethoxy-disubstituted diradicals (1d, Delta E(ST) = -3.6 kcal/mol in C(2) symmetry, and 1e, Delta E(ST) = -3.4 kcal/mol in C(2) symmetry). Spiroconjugation is shown to be responsible for the larger calculated value of absolute value Delta E(ST) in 1g, relative to 1d and 1e. A strong correlation between the calculated values of Delta E(ST) and the computed electronic excitation energies of the singlet diradicals is found for diradicals 1d, 1e, and 1g and for 2,2-difluorocyclopentane-1,3-diyl (1c). A similar correlation between Delta E(ST) and lambda(calcd) is predicted for the corresponding 1,3-diphenylcyclopentane-1,3-diyls 3, and the predicted blue shift in the spectrum of 3g, relative to 3e, has been confirmed by experimental comparisons of the electronic absorption spectra of the annelated derivatives 2c, 2e, and 2g in a glass at 77 K. The wavelength of the first absorption band in the singlet diradicals decreases in the order 2e (lambda(onset) = 650 nm) > 2g (lambda(onset) = 590 nm) > 2c (lambda(onset) = 580 nm). The combination of these computational and experimental results provides a sound basis for reassignment of the first electronic absorption band in singlet diradicals 2c, 2e, and 2g to the excitation of an electron from the HOMO to the LUMO of these 2,2-disubstituted derivatives of cyclopentane-1,3-diyl.     

纳米金催化光度法检测食品中的甲醛

采用分光光度法检测以纳米金催化甲醛与菲林试剂反应生成的氧化亚铜微粒在870nm处的吸收值,建立了一种测定食品中甲醛的新方法．研究了纳米金、菲林试剂用量及反应温度、反应时间、共存离子的影响．在最佳实验条件下,甲醛浓度在1．2-600．0μg／mL范围内与吸光值△A线性关系良好,线性回归方程为△A870nm=0．0014C＋0．0627,相关系数为0．9992,检出限为0．45μg／mL．加标回收率为98．0％～100．3％．该法灵敏度高,选择性好,体系干扰少,用于测定腐竹、精制粉丝、红薯粉丝和方便面中甲醛的含量获得满意的结果．     

Spectrophotometric determination of copper in alloys using naphthazarin

Naphthazarin (5,8-dihydroxy-1,4-naphthoquinone; Naph) is proposed as a chromogenic reagent for the spectrophotometric determination of copper(II). The polynuclear complex has a mole ratio of Cu:Naph=4:6 in a 50% v/v ethanol/water medium containing 0.1 M ammonium acetate and 1.5% (w/v) sodium dodecyl sulfate. The copper-naphthazarin complex shows an absorption maximum at 330 nm with a molar absorptivity of 1.84x10(4) l mol(-1) cm(-1). Beer's law is obeyed up to 4.5 ppm of copper(II). The method was applied for copper determination in alloy samples with satisfactory results.     

1,2-和1,4-萘醌248 nm激光光解的瞬态吸收光谱研究

利用纳秒级激光光解动态吸收光谱装置,研究了1,2-和1,4-萘醌中性水溶液的瞬态吸收光谱.发现1,2-萘醌及1,4-萘醌被光电离后形成的阳离子自由基在380nm均有最大吸收,但1,4-萘醌阳离子自由基在衰变过程中又形成了两种新的活性粒子,它们的最大吸收分别位于410和580nm,分析表明:410nm属于1,4-萘醌脱氢自由基的吸收,而580nm很可能归属由于电子转移而形成的瞬态产物.进一步研究发现,1,2-萘醌在中性水溶液中能被248nm激光单光子电离.     

“On-water” organic synthesis: <Emphasis Type="SmallCaps">l</Emphasis>-proline catalyzed synthesis of pyrimidine-2,4-dione-, benzo[<Emphasis Type="Italic">g</Emphasis>]- and dihydropyrano[2,3-<Emphasis Type="Italic">g</Emphasis>]chromene derivatives in aqueous media

In this work, functionalized pyrimidine-2,4-dione-, benzo[g]-, and dihydropyrano[2,3-g]chromene derivatives have been synthesized via a Michael addition of 2-hydroxy-1,4-naphthoquinone or 2,5-dihydroxy-1,4-benzoquinone to the Knoevenagel condensation product of an aldehyde with Meldrum’s acid, dimedone or barbituric acid in the presence of a catalytic amount of l-proline under refluxing conditions in water in good to excellent yields.     

PHOTOREACTIVATING ENZYME FROM ESCHERICHIA COLI

Abstract— Escherichia coli photoreactivating enzyme (PRE) has been purified in large amounts from an E. coli strain lysogenic for a defective Λ bacteriophage carrying the phr gene. The resulting enzyme has a pH optimum of 7.2 and an ionic strength optimum of 0.18. It consists of an apoprotein and cofactor, both of which are necessary for catalytic activity. The apoprotein has a monomer molecular weight of 35 ,200 and shows stable aggregates under denaturing conditions. The amino acid analysis of the E. coli enzyme is very similar to that of the photoreactivating enzyme from orchid seedlings ( Cattelya aurantiuca ). Both have arginine at the amino terminus. The cofactor, like the holoenzyme, shows absorption, magnetic circular dichroism, and emission properties indicative of an adenine moiety. Although the isolated enzyme has an action spectrum which peaks at about 360 nm, neither the cofactor, apoenzyme nor holoenzyme shows any detectable absorption between 300 and 400 nm.     

Reductively activated nitrous oxide reductase reacts directly with substrate

In the terminal step of bacterial denitrification, N2O is converted to N2 at the mu4-sulfide bridged tetranuclear CuZ center of nitrous oxide reductase. The enzyme can be activated by reduced methyl viologen, with up to a 15-fold increase in specific activity. The reductively activated nitrous oxide reductase from Achromobacter cycloclastes was isolated and characterized by visible absorption and EPR spectroscopy, and both methods showed that the CuZ center can attain a [4Cu(I)] oxidation state. When N2O was added to the activated, reductant-free enzyme, distinct spectral changes were observed, indicating that this state of the enzyme interacts with substrate. This was further supported by the detection of 15N-labeled product in the absence of steady-state turnover conditions. A new absorption band around 970 nm appeared following reaction of activated nitrous oxide reductase with N2O, which may represent a catalytic intermediate state of the enzyme.     

Formation of the early photoproduct lumi-R of cyanobacterial phytochrome cph1 observed by ultrafast mid-infrared spectroscopy

The photoreactions of the Pr ground state of cyanobacterial phytochrome Cph1 from Synechocystis PCC 6803 have been investigated by picosecond time-resolved mid-infrared spectroscopy at ambient temperature. With femtosecond excitation of the Pr state at 640 nm, the photoisomerized Lumi-R product state is generated with kinetics and associated difference spectra indicative of vibrational cooling with tau(1) = 3 ps time constant and excited state decay with tau(1) = 3 ps, tau(2) = 14 ps, and tau(3) = 134 ps time constants. The Lumi-R state is characterized by downshifted absorption of three C=C modes assigned to C(15)=C(16), C(4)=C(6), and a delocalized C=C mode, in addition to the downshifted C(19)=O mode. The Lumi-R minus Pr difference spectrum is indicative of global restructuring of the chromophore on the ultrafast timescale, which is discussed in light of C(15) Z/E photoisomerization in addition to changes near C(5), which could be low bond order torsional angle changes.     

一个新颖三维配位聚合物Pb(1，4-napdc)(DMF)的合成、结构及热稳定性研究

A novel 3D metal-organic coordination polymer Pb(1,4-napdc)(DMF) (1,4-napdc=naphthalene-1,4-dicarboxylate) was synthesized at room temperature using slow vapor diffusion method to grow single crystal that has been analyzed by X-ray diffraction. The crystal belongs to orthorhombic with space group P2₁2₁2₁. The unit cell parameters are as fellows: a=0.701 3(2) nm, b=1.407 6(3) nm, c=1.521 5(4) nm, V=1.501 8(6) nm³ and Z=4. In the crystal structure of Pb(1,4-napdc)(DMF), the square grids constructed with paddle-wheel units of Pb(Ⅱ) and 1,4-napdc links stack over each other to generate infinite 3D network, which has square apertures (1.158×1.158 nm²) along the crystallographic a-axis. The thermal stability of compound was investigated by differential scanning calorimetry and thermogravimetric analysis. CCDC: 293617.     

Chemical, pulse radiolysis and density functional studies of a new, labile 5,6-indolequinone and its semiquinone

The chemical and spectroscopic characterization of 5,6-indolequinones and their semiquinones, key transient intermediates in the oxidative conversion of 5,6-dihydroxyindoles to eumelanin biopolymers, is a most challenging task. In the present paper, we report the characterization of a novel, relatively long-lived 5,6-indolequinone along with its semiquinone using an integrated chemical, pulse radiolytic, and computational approach. The quinone was obtained by oxidation of 5,6-dihydroxy-3-iodoindole (1a) with o-chloranil in cold ethyl acetate or aqueous buffer: it displayed electronic absorption bands around 400 and 600 nm, was reduced to 1a with Na2S2O4, and reacted with o-phenylenediamine to give small amounts of 3-iodo-1H-pyrrolo[2,3-b]phenazine (2). The semiquinone exhibited absorption maxima at 380 nm (sh) and 520 nm and was detected as the initial species produced by pulse radiolytic oxidation of 1a at pH 7.0. DFT investigations indicated the 6-phenoxyl radical and the N-protonated radical anion as the most stable tautomers for the neutral and anion forms of the semiquinone, respectively. Calculated absorption spectra in water gave bands at 350 (sh) and 500 nm for the neutral form and at 310 and 360 (sh) nm for the anion. Disproportionation of the semiquinone with fast second-order kinetics (2k = 1.1 x 1010 M-1 s-1) gave a chromophore with absorption bands resembling those of chemically generated 1a quinone. Computational analysis predicted 1a quinone to exist in vacuo as the quinone-methide tautomer, displaying low energy transitions at 380 and 710 nm, and in water as the o-quinone, with calculated absorption bands around 400 and 820 nm. A strong participation of a p orbital on the iodine atom in the 360-380 nm electronic transitions of the o-quinone and quinone-methide was highlighted. The satisfactory agreement between computational and experimental electronic absorption data would suggest partitioning of 1a quinone between the o-quinone and quinone-methide tautomers depending on the medium.     

Novel Chromogenic Substrate for Bacterial β-Lactamases Based on Cephalosporin Modified with an Epoxy Group

Beta-lactamases are the key enzymes involved in resistance to beta-lactam antibiotics in pathogenic bacteria causing infectious diseases. The search for new inhibitors and the study of the resistance mechanisms require the production of chromogenic substrates for beta-lactamases. A novel cephalosporin derivative with an epoxy functional group named CMPD1 is synthesized. It is shown to be a substrate for TEM type beta-lactamases, which is hydrolyzed to form a colored product. The hydrolysis product has an optical absorption maximum at 450 nm. The difference in the absorption maxima of the substrate and the product is 95 nm, and, therefore, CMPD1 exceeds the previously described substrates, according to this parameter. It has been found that the CMPD1 compound is hydrolyzed only by the TEM type beta-lactamases that lack mutations in the active site. This can be used to study the mechanisms of the catalytic effect of beta-lactamases.     

用手性Rh(Ⅰ)复合物由3-甲基-2-氧代丁酸多相光催化不对称合成L-α-缬氨酸

本文研究了在甲醇和氨水的水溶液里,手性的(+)-diop-Rh(Ⅰ)复合物(diop:2,3-邻-异亚丙基-2,3-二羟基-1,4-双(二苯膦基)丁烷)作为不对称催化剂,粉末半导体CdS、TiO₂、Pt/CdS和Pt/TiO₂为光催化剂,用500WXe灯光照10h,3-甲基-2-氧代丁酸光催化不对称合成L-α缬氨酸已经实现,光学纯度最高达64%e.e.。     

多齿配体2,5-二羟基-1,4-苯醌缩氨基脲 (C_6H_4O_3NNHCONH_2.CHON(CH3_)_2)的合成及晶体结构

报道了含1个溶剂DMF的多齿配体2,5-二羟基-1,4-苯醌缩氨基脲的合成和晶体结构。该化合物(C10N4O5H14)属正交晶系,其空间群为P212121 a=13.221(6),b=17.709(5),c=5.347(4)?, V=1252.0(1)3,Z=4,F(000)=568, Dx=1.434g/cm3,Mr=270.25, m (MoKα)= 1.16 cm-1, 结构由直接法解出,全矩阵最小二乘法修正,最终的偏离因子R=0.0319,wR=0.0748。结构测定表明溶剂DMF分子和2,5-二羟基-1,4苯醌缩氨基脲间形成很强的氢键,后者可看作是两条共轭链的偶合物。     

Two-photon excitation of a phytofluor protein

Phytofluors are highly fluorescent proteins in which the chromophore in a phytochrome is replaced with phycoerythrobilin (PEB), the pigment precursor of the cyanobacterial light harvesting protein phycoerythrin. We examined the fluorescence spectra of the N-terminal region of the cyanobacterial phytochrome 1 from cyanobacterium Synechocystis sp. Pcc 6803 bound to PEB. This protein, Cph1(N514)-PEB, displayed a good two-photon cross-section of 20–30 GM for excitation at 792 nm. This phytofluor also exhibits a high fundamental anisotropy at most practical two-photon excitation (2PE) wavelengths from 700 to 900 nm. Identical lifetimes and correlation times with one and 2PE indicates that the phytofluor is not adversely affected by the intensities needed for 2PE. The one-photon absorption extends well beyond the absorption spectrum and even beyond the emission spectrum to 700 nm. The phytofluor thus appears to be a suitable probe for 2PE and/or cellular imaging.     

Metal‐Free‐Mediated Oxidation Aromatization of 1,4‐Dihydropyridines to Pyridines Using Visible Light and Air

A metal‐free and environmentally friendly aerobic aromatization photosensitized by organic dye eosin Y bis(tetrabutyl ammonium salt) (TBA‐eosinY) has been developed. With the aid of K₂CO₃, the aerobic catalytic system converts 1,4‐dihydropyridines to their corresponding pyridine derivatives efficiently under visible light irradiation (λ=450 nm) at room temperature.     

Asymmetric catalytic allylation of β-diketones or β-ketoesters with allylic ethers using a palladium-DIOP catalyst: A mechanistic study

Allylation of β-diketone, β-ketoesters and methine active hydrogen compounds by allyl phenyl ethers or allyl esters with palladium/phosphine catalytic systems has been studied, and a mechanism is suggested. The use of DIOP (2,3-O-isopropylidene-2,3-dihydroxy-1,4-bis(diphenylphosphino)butane) as chiral phosphine ligand produces optically active allylated compounds with up to 10% e.e.     

4,4′-Dihydroxy-3,3′,5,5′-tetramethoxyazodioxybenzene: an unexpected dimer formed during hydroxylamine extractions of wheat flour

Neutral hydroxylamine extracts of wheat contained a product that was colourless at pH<5 (λ_max 340 nm) and yellow at pH>9 (λ_max 400 nm). ESI-MS showed a major ion m/z 184.0 and a possible parent ion m/z 367.2 (MH⁺) suggesting that the product resulted from the reaction of 2,6-dimethoxy-p-quinone with hydroxylamine. However, mass spectral and other spectroscopic data indicated that the compound was neither of the 2,6-dimethoxy-p-quinone oximes. A product with identical absorbance, mass spectrum, electrophoretic mobility and HPLC retention time as the pigment from hydroxylamine extracts of flour was observed amongst the reaction products of hydroxylamine and 1,4-dihydroxy-2,6-dimethoxybenzene. The structure of this product was identified by NMR, 2D NMR and IR as 4,4′-dihydroxy-3,3′,5,5′-tetramethoxyazodioxybenzene.     

一种新型的紫外吸收剂——二苯甲酰甲烷的光谱特性及光稳定能力的初步研究

本文研究了一种新型的紫外吸收剂——二苯甲酰甲烷的光谱特性。实验结果表明,在基态,它存在酮式-烯醇式的热化学平衡,且烯醇式形成六元环分子内氢键。在顺-1,4-聚丁二烯溶液光降解的比较实验中,二苯甲酰甲烷的光稳定能力和邻羟基-对正辛氧基二苯甲酮相近,并对光稳定行为的可能机理进行了研究。     

检测痕量纳米金的纳米催化光度法

在EDTA—NaOH介质中,金纳米微粒对盐酸联氨还原硫酸铜生成铜微粒这一慢反应具有较强的催化作用．铜微粒在750nm处产生一个吸收峰．随着纳米金浓度的增大,750nm处的吸光强度线性增大．对于粒径为10、30、50nm的纳米金,其线性范围、回归方程、检出限分别为0．12～1．68、0．36～2．80、1．00～5．00nmol／L,△A750nm=0．3205CAu＋0．0076、△A750nm=0．2201CAu＋0．0056、△A750nm=0．1150CAu＋0．0066,0．05、0．20、0．50nmol／L Au．分别对0．50、1．00nmol／L纳米金（d=10nm）平行测定10次,求得其相对标准偏差分别为4．2％、3．5％．