硫胺的燐光研究

硫胺无燐光，在碱中氧化后才产生燐光。 λ激发＝３００ｎｍ，λ发射＝４１０ ｎｍ，燐光寿命为１．３ｓ．硫胺燐光受溶剂极性影响甚小，１５种金属离子中只有铜离子可增强流胺燐光，寿命变短为 ０． ９４ ｓ，但不影响燐光光谱。在开始３０ｍｉｎ内有ＣｕＳＯ４的作用较快，以后逐渐变慢，到 ４ｈ后达平衡与不用ＣｕＳＯ４的燐光相近，启示ＣｕＳＯ４具有催化硫胺的氧化作用，建立了硫胺的燐光分析方法，用于两种维生素Ｂ１片的分析，分别为９８．５％与９８％。     

A chromophore-supported structural and functional model of dinuclear copper enzymes,for facilitating mechanism of action studies

Type III dicopper centres are the heart of the reactive sites of enzymes that catalyze the oxidation of catechols. Numerous synthetic model complexes have been prepared to uncover the fundamental chemistry involved in these processes, but progress is still lagging much behind that for heme enzymes. One reason is that the latter gain very much from the informative spectroscopic features of their porphyrin-based metal-chelating ligand. We now introduce sapphyrin-chelated dicopper complexes and show that they may be isolated in different oxidation states and coordination geometries, with distinctive colors and electronic spectra due to the heme-like ligands. The dicopper(i) complex 1-Cu2 was characterized by ¹H and ¹⁹F NMR spectroscopy of the metal-chelating sapphyrin, the oxygenated dicopper(ii) complex 1-Cu2O2 by EPR, and crystallographic data was obtained for the tetracopper(ii)-bis-sapphyrin complex [1-Cu2O2]2. This uncovered a non-heme [Cu₄(OH)₄]⁴⁻ cluster, held together with the aid of two sapphyrin ligands, with structural features reminiscent of those of catechol oxidase. Biomimetic activity was demonstrated by the 1-Cu2O2 catalyzed aerobic oxidation of catechol to quinone; the sapphyrin ligand aided very much in gaining information about reactive intermediates and the rate-limiting step of the reaction.

Di-copper chelation by sapphyrin facilitates reaction mechanism investigations and characterization of reactive intermediates regarding biomimetic catechol oxidation.     

Research on synthesis and action mechanism of polycarboxylate superplasticizer

The relationship between the structure and performance of polycarboxylate superplasticizer was analyzed. The respective functions of the structure units of the main and branched chains were discussed. The progress of synthesis and molecular structure design and synthesis of polycarboxylate superplasticizer were reviewed according to the difference in the structure unit of the main chain. Results indicated that their performance is related to the structure unit of the main and branched chains, as well as the position and species of functional groups. The polycarboxylate superplasticizer, which had suitable graft and block polymers of polyethylene glycol or polyoxyethylene, and a suitable sulfonic group, had small slump loss besides high water-reducing performance. On the other hand, the hydroxyl group at the end of the chain causes gelation easily. On the basis of the items mentioned above, as well as the source and cost of raw materials, esterification of polyethylene glycol and acrylic acid were first adopted using para-toluene sulfonic acid as catalyst, then polymerized with sodium sulfonate methacrylate. A certain amount of acrylic was added in order to regulate both the polymerization degree of the main chain and the ratio of carboxyl and sulfonic groups in the branched chain. As a result, the high performance superplasticizer has been synthesized (was obtained). The divergence of the cement plasma is about 200 mm when the addition amount of superplasticizer is 0.16%–0.20% of cement weight, and the ratio of the water and cement is 0.29. Translated from Journal of Wuhan Uniwersity of Technology, 2006, 28(9): 18–20 [译自: 武汉理工大学学报]     

Inferring the chemical mechanism from structures of enzymes

Chemistry has once again embraced the study of enzyme mechanism as a core discipline. Chemists are uniquely able to contribute to the analysis of enzymes through their understanding of the reactivity of atoms. In this tutorial review for the Corday-Morgan medal, I will concentrate on the work from my lab over the past six years. I discuss enzymes which transform carbohydrates and incorporate halogens. The tutorial review will emphasise the strengths and limitations of structural biology as a means to deducing the chemical mechanism.     

Daptomycin structure and mechanism of action revealed

Daptomycin kills otherwise antibiotic-resistant gram-positive pathogens and is the first lipopeptide antibiotic to reach the clinic. Elucidation of its 3D structure and mechanism of action, reported in this issue of Chemistry & Biology, will facilitate the design and engineering of new, potentially life-saving antibiotics.     

钒(Ⅳ)-硫胺素荧光光度法的研究与应用

利用钒(Ⅳ)作为氧化剂将硫胺素氧化成硫胺荧,建立了一种测量痕量硫胺素的荧光分光光度法,并将该方法应用于药物中硫胺素的测定.该方法测定硫胺素的线性范围为0.8～240.0μg/L;检出限为0.45μg/L;对于40.0μg/L的硫胺素测定10次,其相对标准偏差为1.5%.     

Modeling the action of drugs on cellular enzymes by means of optimal control techniques

In this paper we apply Optimal Control techniques, based on the solution of Hamilton–Jacobi–Bellman equations, to simulate the action of a drug on some sub-networks of an intracellular signal transduction network. The cost functional to be minimized is chosen in order to take into account, in the models, different factors, like the toxicity of some reactants, the drug action and the costs of the drug itself. Numerical results are also shown, to test the model.     

Enzymatic degradation of chitosan films under the action of nonspecific enzymes

The features of the enzymatic degradation process mediated by the nonspecific enzymes collagenase and Lyrase are discussed for chitosan films prepared from acetic acid solutions. It was shown that variations in the acid concentration in the initial solution have a substantial effect on both the structure of chitosan in the film formed and the degree of degradation of film samples.     

Conservation of mechanism in three chorismate-utilizing enzymes

Chorismate is the end-product of the shikimate pathway for biosynthesis of carbocyclic aromatic compounds in plants, bacteria, fungi, and some parasites. Anthranilate synthase (AS), 4-amino-4-deoxychorismate synthase (ADCS), and isochorismate synthase (IS) are homologous enzymes that carry out the initial transformations on chorismate in the biosynthesis of tryptophan, p-aminobenzoate, and enterobactin, respectively, and are expected to share a common mechanism. Poor binding to ADCS of two potential transition state analogues for addition of a nucleophile to C6 of chorismate implies that it, like AS and IS, initiates reaction by addition of a nucleophile to C2. Molecular modeling based on the X-ray structures of AS and ADCS suggests that the active site residue K274 is the nucleophile employed by ADCS to initiate the reaction, forming a covalent intermediate. The K274A and K274R mutants were shown to have 265- and 640-fold reduced k(cat) values when PabA (the cognate amidotransferase) + glutamine are used as the nitrogen source. Under conditions of saturating chorismate and NH(4)(+), ADCS and the K274A mutant have identical k(cat) values, suggesting the participation of NH(4)(+) as a rescue agent. Such participation was confirmed by the buildup of 2-amino-2-deoxyisochorismate in the reactions of the K274A mutant but not ADCS, when either NH(4)(+) or PabA + glutamine is used as the nitrogen source. Additionally, the inclusion of ethylamine in the reactions of K274A yields the N-ethyl derivative of 2-amino-2-deoxyisochorismate. A unifying mechanism for AS, ADCS, and IS entailing nucleophile addition to C2 of chorismate in an S(N)2' ' process is proposed.     

Kinetic study on the mechanism of inhibition of trypsin and trypsin-like enzymes by p-guanidinobenzoate ester

It was found that [4-(2-succinimidoethylthio)phenyl 4-guanidinobenzoate]methanesulfonate (E-3123) inhibits trypsin, thrombin and kallikrein, and its inhibitory activity is most potent toward trypsin. The interactions of these enzymes with E-3123 were studied mainly by using stopped-flow spectrophotometry. E-3123 behaved as a quasi-substrate of the enzymes and the inhibitory property was due to the efficient production of the stable acyl-enzyme. The acylation process with trypsin was exceedingly effective, and the resulting acyl-enzyme was the most stable among the three enzymes tested. This observation affords a rational basis for explaining the action of E-3123, which is a transient inhibitor most active toward trypsin.     

Enzymes动能化的树形高分子: 蛋白和酶的人工模型

含有官能机制特殊树枝状聚合物可作为生物有机分子, 如蛋白质及酵素的模型。本文将介绍这类型官能性树枝聚合物的制备方法。     

Theoretical study on the mechanism of the oxygen activation process in cysteine dioxygenase enzymes

Cysteine dioxygenase (CDO) is a vital enzyme for human health involved in the biodegradation of toxic cysteine and thereby regulation of the cysteine concentration in the body. The enzyme belongs to the group of nonheme iron dioxygenases and utilizes molecular oxygen to transfer two oxygen atoms to cysteinate to form cysteine sulfinic acid products. The mechanism for this reaction is currently disputed, with crystallographic studies implicating a persulfenate intermediate in the catalytic cycle. To resolve the dispute we have performed quantum mechanics/molecular mechanics (QM/MM) calculations on substrate activation by CDO enzymes using an enzyme monomer and a large QM active region. We find a stepwise mechanism, whereby the distal oxygen atom of the iron(II)-superoxo complex attacks the sulfur atom of cysteinate to form a ring structure, followed by dioxygen bond breaking and the formation of a sulfoxide bound to an iron(IV)-oxo complex. A sulfoxide rotation precedes the second oxygen atom transfer to the substrate to give cysteine sulfinic acid products. The reaction takes place on several low-lying spin-state surfaces via multistate reactivity patterns. It starts in the singlet ground state of the iron(II)-superoxo reactant and then proceeds mainly on the quintet and triplet surfaces. The initial and rate-determining attack of the superoxo group on the cysteinate sulfur atom involves a spin-state crossing from singlet to quintet. We have also investigated an alternative mechanism via a persulfenate intermediate, with a realignment of hydrogen bonding interactions in the substrate binding pocket. However, this alternative mechanism of proximal oxygen atom attack on the sulfur atom of cysteinate is computed to be a high-energy pathway, and therefore, the persulfenate intermediate is unlikely to participate in the catalytic cycle of CDO enzymes.     

The role of C-centered radicals on the mechanism of action of artemisinin

Artemisinin is a sesquiterpene lactone with an endoperoxide function that is essential for its antimalarial activity. Endoperoxides are supposed to act on heme leading to the reduction of the peroxide bond and production of radicals, which can be responsible for killing the parasite. The geometries of artemisinin, radical anions and neutral species generated by rearrangement after reduction of the peroxide bond were fully optimized with the AM1 and PM3 semi-empirical methods, in order to characterize the intermediates formed during the process. Among the radicals calculated along the pathway for reductive decomposition of artemisinin, the secondary radical centered on carbon C₄ has the highest stability when compared to other radicals that can be achieved either by hydrogen migration to the original O-centered radical or by homolytic break of C–C bond. This suggests that the C₄-centered radical may be the species responsible for killing the parasite, as has been indicated previously in experimental studies.