Glycosyl fluorides in enzymatic reactions

Glycosyl fluorides have considerable importance as substrates and inhibitors in enzymatic reactions. Their good combination of stability and reactivity has enabled their use as glycosyl donors with a variety of carbohydrate processing enzymes. Moreover, the installation of fluorine elsewhere on the carbohydrate scaffold commonly modifies the properties of the glycosyl fluoride such that the resultant compounds act as slow substrates or even inhibitors of enzyme action. This review covers the use of glycosyl fluorides as substrates for wild-type and mutant glycosidases and other enzymes that catalyze glycosyl transfer. The use of substituted glycosyl fluorides as inhibitors of enzymes that catalyze glycosyl transfer and as tools for investigation of their mechanism is discussed, including the labeling of active site residues. Synthetic applications in which glycosyl fluorides are used as glycosyl donors in enzymatic transglycosylation reactions for the synthesis of oligo- and polysaccharides are then covered, including the use of mutant glycosidases, the so-called glycosynthases, which are able to catalyze the formation of glycosides without competing hydrolysis. Finally, a short overview of the use of glycosyl fluorides as substrates and inhibitors of phosphorylases and phosphoglucomutase is given.     

2-Deoxy-2-fluoro-D-glycosyl fluorides. A new class of specific mechanism-based glycosidase inhibitors

Mechanism-based glycosidase inhibitors are of considerable use in studies of enzyme mechanism, in studies of glycoprotein processing, and possibly therapeutically in control of sugar uptake. This paper describes a new general approach to mechanism-based inactivation of glycosidases which involves trapping a covalent glycosyl enzyme intermediate. This is achieved by use of 2-deoxy-2-fluoro-D-glycosyl fluorides, for which the rate of hydrolysis of the fluoroglycosyl enzyme intermediate is extremely slow, resulting in accumulation of the intermediate. Eleven different glycosidases were tested with their corresponding 2-deoxy-2-fluoro-D-glycosyl fluorides. Eight of the eleven were inactivated, four of them according to pseudo first-order kinetics and four according to a more complex kinetic scheme. The specificity of these inhibitors was investigated by assaying for inhibition of one enzyme with four different 2-deoxy-2-fluoro-D-glycosyl fluorides. Large differences in inactivation rate were observed which paralleled previously observed substrate specificities.     

氟代糖在糖苷酶研究中的应用

近年来,氟代糖应用于糖苷酶反应研究,显示出越来越重要的作用。氟代糖可以作为糖苷酶及其突变酶的水解底物研究酶学性质;氟代糖抑制剂可以标记糖苷酶催化中心,鉴定亲核体氨基酸。尤为重要的是,氟代糖可作为糖苷酶的糖基供体来合成糖类。糖苷酶突变后,可生成糖苷合成酶和硫代糖苷合成酶,可以用与正常底物构型相反的氟代糖作为糖基供体高效合成糖类,收率一般为60%~90%,有的可达100%。糖苷酶及其突变酶以氟代糖为底物高效合成糖类的研究,必将促进生物学、糖生物学和纳米生物材料的发展。     

Impact of fluorides on the removal of heavy metals from an electroplating effluent using a flocculent brewer’s yeast strain of Saccharomyces cerevisiae

Abstract

Besides several toxic heavy metals, electroplating effluents can have in solution different cations and anions, which may influence heavy metals removal by the biomass. Among them, fluorides are commonly used in the electroplating industries and thus can be found in the respective wastewaters. In the present work, the effect of the presence of fluorides in the efficiency of chromium(III), copper(II) and nickel(II) removal, from an effluent, by heat-inactivated cells of a brewing flocculent strain of Saccharomyces cerevisiae was evaluated. The presence of fluorides severely decreased (>60%) the removal of chromium(III) by yeast biomass. This effect impaired the effective treatment of the effluent according to the US Environmental Protection Agency and the Portuguese law; conversely, a higher removal of copper(II) and nickel(II) was observed. This behaviour can be understood by metal speciation. In the presence of fluorides, chromium(III) was mainly complexed, becoming unavailable for yeast accumulation; this effect decreased the efficiency of chromium(III) removal. Thus, in the presence of fluorides, less chromium(III) is associated with biomass and consequently more yeast binding sites remain available for the uptake of other metals present in solution. This fact explains the increase of copper(II) and nickel(II) removal in the presence of fluorides.     

Inhibition of lipoprotein lipase by alkanesulfonyl fluorides

A number of alkanesulfonyl halides (chlorides and fluorides) and esters were synthesized and their effect on the activity of lipoprotein lipase (LPL) was studied. Sulfonyl fluorides proved to be efficient inhibitors of LPL when the enzyme was incubated with a 10-fold molar excess of the inhibitors in a buffer containing bile salts (deoxycholate). Hexadecane- and dodecanesulfonyl fluorides caused 50% inhibition of LPL activity at concentrations of 10 to 20 μM.     

Humus status of different soils affected by pollution with fluorides from aluminum production in the Baikal region

The effect of pollution by fluorides from aluminum production on the humus status in different types of arable soils was studied in field experiments. They included modeling of a high pollution level by addition of NaF prevailing in industrial air emissions. The negative effect of fluorides on the humus status, related to an increase in the mobility of humic substances, was more pronounced in gray forest soil than in sod-meadow soil. This depended on the soil physicochemical properties determining the buffer capacity toward NaF and the content of water-soluble fluorides.     

Salivary Fluoride Concentrations and Fluoride Ingestion Following Application of Preparations Containing High Concentration of Fluoride

The aim of the study was to assess salivary fluoride concentrations and ingestion of fluorides after five different techniques of topical fluoride treatment. Ten volunteers received applications of fluoride gel, toothpaste, and foam. Fluoride concentrations were determined using ion-selective fluoride electrode in the samples of unstimulated saliva before and after procedures. The amounts of fluoride applied, recovered from the mouth, and retained in the mouth were calculated for each treatment. It was proved that fluoride ingestion following tray application of fluoride foam was significantly lower than ingestion following tray applications of fluoride gel (p < 0.01). The use of limited amounts of fluorides on a tray resulted in similar fluoride retention and lower ingestion comparing to the method which involves a large portion of fluorides followed by rinsing. Tooth brushing resulted in high salivary retention rates per amount of fluorides used in the procedure. These data provide initial concept about the possible advantages of some methods of topical fluoride application over others.     

Synthesis and biological evaluation of novel irreversible serine protease inhibitors using amino acid based sulfonyl fluorides as an electrophilic trap

We have designed and synthesized novel irreversible serine protease inhibitors containing aliphatic sulfonyl fluorides as an electrophilic trap. These substituted taurine sulfonyl fluorides derived from taurine or protected amino acids were conveniently synthesized from β-aminoethanesulfonyl chlorides using KF/18-crown-6 or from β-aminoethanesulfonates using DAST. Their potency of irreversible inhibition of serine proteases is described in different enzyme assays using chymotrypsin leading to binding affinities up to 22 μM.     

兰州一个铝厂氟污染对植物的影响

木文是以兰州一个铝厂为中心,沿北面、西北面、东南面和南面四个方向设置十八个测点。采集空气样品235个,土壤样品127个,植物样品362个。这些样品的测定结果,说明空气和土壤的污染主要在2—4公里的半径范围内,下风向可能到达7.5公里。由于植物从空气中吸收氟,并加以积累,使叶片含量达数百ppm,最高达2534ppm,构成氟的伤害。这种伤害最早(5月间)出现在离铝厂最近测点的桃树(Prunus persica)、杏树(P．armeniaca)上,以后逐渐扩大到稍远的其他地方和别的植物,使葱、蒜失去食用价值,小麦发育不良,叶面积缩小,有效分蘖下降,穗长缩短,小穗数减少,千粒重从50.1克下降为37.6克,造成粮食减产。根据铝厂周围地区植物、土壤和空气氟的污染程度,可分为四种类型：清洁区、轻污染区、污染区和重污染区。     

The hydrolysis of glycosyl fluorides by glycosidases. Determination of the anomeric configuration of the products of glycosidase action

The enzymic hydrolysis of glycosyl fluorides is conveniently followed by using a pH-stat. Reactions involving glucosyl or galactosyl fluorides can also be followed by using glucose oxidase or galactose oxidase respectively. The pH-stat allows the rapid assay of intestinal alpha-glucosidase in crude homogenates. Use of glycosyl fluorides as substrates for glycosidases facilitates the polarimetric or g.l.c. determination of the anomeric nature of the initial product of hydrolysis. Hydrolysis by fungal amyloglucosidase proceeds with inversion of configuration whereas that by yeast and rat intestinal alpha-glucosidase, coffee-bean alpha-galactosidase and almond emulsin beta-glucosidase proceeds with retention of configuration. beta-d-Glucopyranosyl azide was not a detectable substrate for almond emulsin beta-d-glucosidase.     

Chemical Origins of Electrochemical Overpotential in Surface‐Conversion Nanocomposite Cathodes

A new branch of promising nanocomposite cathode materials for rechargeable batteries based on non‐intercalation materials has been recently discovered. However, all the nanocomposite cathodes reported thus far suffer from a large overpotential in the first charge, which hinders the activation and lowers the energy efficiency. Here, a series of model nanocomposites consisting of MnO and various metal fluorides (LiF, NaF, KF, RbF, CsF, MgF₂, CaF₂, and AlF₃) to identify the key parameters affecting the activation and overpotential in the first charge are evaluated. It is demonstrated that the F 1s binding energy of the metal fluorides is a plausible indicator of the overpotential in the first charge as well as the subsequent reversible discharge capacity. The stability of the cation in the electrolyte and its solvation nature are also shown to affect the overall activation process. Finally, it is proposed that appropriate tuning of the binding energy of metal fluorides (e.g., by forming solid solutions such as LiCsF₂) is a feasible approach to reduce the overpotential and increase the reversible capacity. The findings broaden the current understanding of surface‐conversion nanocomposite chemistries, thus providing guidelines for the design of nanomixture cathode materials for rechargeable batteries.     

Synthesis of Fmoc-amino acid fluorides via DAST, an alternative fluorinating agent

Summary DAST [(diethylamino)sulfur trifluoride] has shown to be an excellent alternative fluorinating agent for synthesis of Fmoc-amino acid fluorides. The use of DAST is advantageous relative to other fluorinating agents, such as cyanuric fluoride, since handling and isolation of the acid fluorides are facile.     

Methods of synthesis of glycosyl fluorides

In recent years glycosyl fluorides have been utilized as versatile sugar donors in the synthesis of natural products and carbohydrates. This paper provides an update on the advances made in the preparation of glycosyl fluorides during the last decade (1988-1998).     

Colorimetric detection of in situ metal acetates and fluorides by a bipyridyl‐linked Schiff base

Here, we present a new bipyridyl moiety linked Schiff base (bipy‐1) that is well characterized using spectroscopic techniques. Colorimetric and UV‐vis titrations were used to study the photophysical properties of bipy‐1 in the presence of various tetrabutyl ammonium salt of anions and metal salts containing different counter cations. bipy‐1 showed selective recognition of dimethyl sulphoxide solution of tetrabutyl ammonium salt of F^? ion accompanied with a UV‐vis band at 529 nm and interesting binding of aqueous Co, Ni, and Cu acetates/fluorides, as confirmed by distinct color changes from fluorescent green to pink or orange and a strong band around 480–510 nm in the UV‐vis spectrum. However, in the presence of Co, Ni, and Cu countercations, any form of metal acetate/fluorides was found to be able to respond to similar color changes from fluorescent green to pink or orange, showing a band around 480–510 nm. This type of output clearly indicates that the in situ formation of Co, Ni, and Cu acetates/fluorides also coordinates with bipyridyl nitrogen atoms. Copyright © 2014 John Wiley & Sons, Ltd.     

Changes in the content of zinc and fluoride during growth of the femur in chicken

Zinc and fluorides are capable of modifying the process of bone formation and mineralization. Statistically significant differences have been revealed in the content of zinc and fluorides between structures of the femur in chicken. The content of zinc in compact bone remained constant during the first 50 d of life. Lower and less stable contents were found in spongy bone and bone marrow. The content of fluorides in compact bone was higher than in spongy bone. The lowest concentrations of zinc and fluorides were found in articular cartilage and were further reduced at the end of observation. Correlations revealed between the content of zinc and fluorides point to structural and functional relationships between these elements in various parts of the bone.     

The influence of fluorides on mouse sperm capacitation

Increasing infertility, due to pathological changes on sperm, has become a serious issue. Eco-toxicological effect of rising concentration of fluorides can be enhanced in the presence of aluminium ions by forming fluorometallic complexes, analogues of phosphate groups that interfere with the activity of G-proteins and P-type ATPases, which are part of several signalling pathways during sperm maturation. In order for sperm to gain fertilizing ability, they must undergo in the female reproductive tract, capacitation that includes tyrosine phosphorylation and consequent actin polymerization. The present paper reports the findings of 3-month oral toxicity in mice of fluorides at the concentrations 0, 1, 10, and 100ppm and their synergic action with aluminium at dose of 10ppm. There were no mortalities, clinical signs of discomfort or body weight loss during the experiment. The analysis revealed, for the concentrations of 10 and 100ppm, abnormalities of spermatogenesis and ability of epididymal spermatozoa to capacitate in vitro, as the result of decreased sperm head tyrosine phosphorylation and actin polymerization. The enhancing overload caused by fluorides represents a potential factor, having an impact on function of sperm, hence contributing to a growing infertility in the human population.     

Assessment of the Bioaccessibility of Trace Elements in Cat’s Claw Teas by In Vitro Simulated Gastrointestinal Digestion Using FAAS

Beer is a widely consumed drink throughout the world, and because its manufacture involves the use of water, beer can be, in some cases, a source of fluorides. For this reason, the objective of this study was to determine the concentration of fluorides in 50 samples of beers from different sources sold in two different types of container (aluminum can and glass bottle). The possible significant differences between the different types of packaging and the intake of fluoride from the consumption of these beers were evaluated. The concentration of fluoride in beers has been determined using the potentiometric method of fluoride determination by standard addition. The concentration of fluoride ranged between 0.06 and 1.77 mg/L. In general, the concentration was below 1 mg/L, except for three beer samples from Ireland and the USA, whose concentration was over 1.5 mg/L. No significant differences were found between the types of packaging. The contribution of fluoride to the diet from beer consumption is not high (<27%); however, it is necessary to warn consumers whenever they are in areas of high concentrations of fluoride in the water supply.     

Convenient and efficient synthesis of Boc-/Z-/Fmoc-beta-amino acids employing N-protected alpha-amino acid fluorides.

A new and efficient method for the synthesis of N(alpha)-Fmoc-/Boc-/Z-beta-amino acids using the two-step Arndt-Eistert approach is described. Fmoc-/Boc-/Z-alpha-Amino acid fluorides were used for the acylation of diazomethane synthesizing Fmoc-/Boc-/Z-alpha-aminodiazoketones as crystalline solids with good yield and purity. They were then converted to the corresponding beta-amino acids using PhCOOAg/dioxane/H2O.     

Glycosylation with activated sugars using glycosyltransferases and transglycosidases

The growing recognition of the roles of carbohydrates in fundamental biological processes and their potential application as functional foods and new therapeutics have generated a requirement for the general availability of larger amounts of varying carbohydrate structures.

Thus the synthesis of oligo-, polysaccharides and glycosylated substances/products represents a major challenge.

Activated sugars are key substrates for synthesis and glycosylation. Nucleotide activated sugars are natural tools for highly selective synthesis, providing complex polysaccharides, glycopeptides, glycolipids etc. However their high cost and availability limit their application. Sucrose acts as an activated substrate for a range of sucrase enzymes elaborating natural polysacchrides of the glucan and fructan type, which also serve for the synthesis and technical production of different oligosaccharides. Sucrose analogues have been shown to extend the range of oligosaccharide synthesis making new structures available incorporating further monosaccharides, such as mannose, galactose, xylose, rhamnose and fucose. A short overview of the use of glycosyl phosphates and glycosyl fluorides as substrates is also given.     

Ethenesulfonyl fluoride derivatives as telomerase inhibitors: structure-based design,SAR, and anticancer evaluation in vitro

Based on our previous docking model, in order to carry out more rational drug design, totally 82 vinyl sulfonyl fluorides, including some 2-(hetero)arylethenesulfonyl fluorides and 1,3-dienylsulfonyl fluorides derivatives as potential human telomerase inhibitors were designed and synthesised. The in vitro anticancer activity assay showed that compound 57 (1E,3E)-4-(4-((E)-2-(fluorosulfonyl)vinyl)phenyl)buta-1,3-diene-1-sulfonyl fluoride exhibited high activity against A375 and MDA-MB-231 cell lines with IC₅₀ 1.58 and 3.22?µM, but it manifested obvious un-toxic effect against GES-1 and L-02 with IC₅₀ with IC₅₀ values less than 2.00?mM. By the modified TRAP assay, some compounds including 57 exhibited potent inhibitory activities against telomerase with IC₅₀ values of 0.71–0.97?µM.