Preparation of glucose oxidase immobilized in different carriers using radiation polymerization

Glucose oxidase (EC 1.1.3.4) was immobilized on different polymeric materials using different immobilization techniques (entrapping by γ‐irradiation, and covalent binding using epichlorohydrin). Studies were carried out to increase the thermal stability of glucose oxidase (GOD) for different applications. The activity and stability of the resulting biopolymers have been compared with those of free GOD. The effect of different polyvinyl alcohol/polyacrylamide (PVA/PAAm) compositions of the copolymer carrier on the enzymatic activity of the immobilized GOD was studied. The maximum enzymatic activity was obtained with the composition ratio of PVA/PAAm of 60:40. The behaviour of the free and immobilized enzyme was analysed as a function of pH. A broadening in the pH profile (5.5–8) was observed for immobilized preparations. The activity and stability of the resulting biopolymers produced by immobilization of GOD onto different carriers have been compared, in both aqueous and organic media, with those of the free GOD. The enzyme's tolerance toward both heat and organic solvent was enhanced by immobilization onto polymers. The addition of different concentrations of organic solvents (10–50%, v/v) to the enzyme at higher temperature (60 °C) was found to stabilize the enzyme molecule. The strongest stabilizing effect on the enzymatic activity was achieved at a concentration of 10%. Copyright © 2005 Society of Chemical Industry     

Electrochemical behavior of metallic materials used in seawater—interactions between glucose oxidase and passive layers

Six materials (stainless steel, nickel-base alloy, titanium, chromium, nickel and admiralty brass) are tested in chemical and biochemical synthetic seawaters. The biochemical seawater contains enzymes catalyzing oxidation of glucose (glucose oxidase), simulating the action of natural biofilms. The evolutions of free corrosion potential (Ecor) versus time, and of cathodic and anodic reactions are compared with those obtained in natural seawater. Then, electrochemical behavior is related to semi-conducting properties of passive films. When glucose oxidase is present, increase of Ecor versus time is only reported for materials presenting a n-type semi-conductor passive film, and whose cathodic reaction current is increased. On the contrary, when passive layers are p-type semi-conductors, cathodic and anodic reactions are increased, and lead to a global Ecor constant with time of immersion. It appears that interaction between bacteria, medium and materials includes evolution of semi-conducting properties of passive layers.     

Antioxidant activity of anacardic acids

Anacardic acids, 6-pentadec(en)ylsalicylic acids isolated from the cashew Anacardium occidentale L. (Anacardiaceae) nut and apple, were found to possess preventive antioxidant activity while salicylic acid did not show this activity. These anacardic acids prevent generation of superoxide radicals by inhibiting xanthine oxidase (EC 1.1.3.22, Grade IV) without radical-scavenging activity. Notably, the inhibition kinetics of anacardic acids do not follow hyperbolic dependence of enzyme inhibition on inhibitor contents (Michaelis–Menten equation) but follow the Hill equation instead. Anacardic acid (C_15:1) inhibited the soybean lipoxygenase-1 (EC 1.13.11.12, Type 1) catalyzed oxidation of linoleic acid with an IC₅₀ of 6.8 μM. The inhibition is a slow and reversible reaction without residual enzyme activity. The inhibition kinetics indicate that anacardic acid (C_15:1) is a competitive inhibitor and the inhibition constant, K_I, was 2.8 μM. Anacardic acids act as antioxidants in a variety ways, including inhibition of various prooxidant enzymes involved in the production of the reactive oxygen species and chelate divalent metal ions such as Fe²⁺ or Cu²⁺, but do not quench reactive oxygen species. The C₁₅-alkenyl side chain is largely associated with the activity.     

Regulation of Neutrophil Functions by Hv1/VSOP Voltage-Gated Proton Channels

The voltage-gated proton channel, Hv1, also termed VSOP, was discovered in 2006. It has long been suggested that proton transport through voltage-gated proton channels regulate reactive oxygen species (ROS) production in phagocytes by counteracting the charge imbalance caused by the activation of NADPH oxidase. Discovery of Hv1/VSOP not only confirmed this process in phagocytes, but also led to the elucidation of novel functions in phagocytes. The compensation of charge by Hv1/VSOP sustains ROS production and is also crucial for promoting Ca²⁺ influx at the plasma membrane. In addition, proton extrusion into neutrophil phagosomes by Hv1/VSOP is necessary to maintain neutral phagosomal pH for the effective killing of bacteria. Contrary to the function of Hv1/VSOP as a positive regulator for ROS generation, it has been revealed that Hv1/VSOP also acts to inhibit ROS production in neutrophils. Hv1/VSOP inhibits hypochlorous acid production by regulating degranulation, leading to reduced inflammation upon fungal infection, and suppresses the activation of extracellular signal-regulated kinase (ERK) signaling by inhibiting ROS production. Thus, Hv1/VSOP is a two-way player regulating ROS production. Here, we review the functions of Hv1/VSOP in neutrophils and discuss future perspectives.     

The haustorium and the chemistry of host recognition in parasitic angiosperms

Two parasitic angiosperms,Agalinis purpurea (Scrophulariaceae) andStriga asiatica (Scrophulariaceae), are compared as to the chemical recognition events involved in host selection.Agalinis is a hemiparasite which can mature to seed-set without a host, whereasStriga is a holoparasite and survives for only a very limited time without a host. Both parasites, however, attach to a host through a specialized organ known as the haustorium and regulate the development of this organ through the recognition of chemical factors from host plants. We now describe the discovery of 2,6-dimethoxy-p-benzoquinone (2,6-DMBQ) as an haustoria-inducing principle fromSorghum root extracts. Our investigation of this compound has led us to suggest that one level of host recognition in these parasitic plants is mediated through their enzymatic digestion of the host root surface. Degradation of surface components liberates quinonoid compounds, such as 2,6-DMBQ, which in turn trigger haustorial development.     

Activation of plant foliar oxidases by insect feeding reduces nutritive quality of foliage for noctuid herbivores

The foliage and fruit of the tomato plantLycopersicon esculentum contains polyphenol oxidases (PPO) and peroxidases (POD) that are compartmentally separated from orthodihydroxyphenolic substrates in situ. However, when leaf tissue is damaged by insect feeding, the enzyme and phenolic substrates come in contact, resulting in the rapid oxidation of phenolics to orthoquinones. When the tomato fruitwormHeliothis zea or the beet army-wormSpodoptera exigua feed on tomato foliage, a substantial amount of the ingested chlorogenic acid is oxidized to chlorogenoquinone by PPO in the insect gut. Additionally, the digestive enzymes of the fruitworm have the potential to further activate foliar oxidase activity in the gut. Chlorogenoquinone is a highly reactive electrophilic molecule that readily binds cova-lently to nucleophilic groups of amino acids and proteins. In particular, the —SH and —NH₂ groups of amino acids are susceptible to binding or alkylation. In experiments with tomato foliage, the relative growth rate of the fruitworm was negatively correlated with PPO activity. As the tomato plant matures, foliar PPO activity may increase nearly 10-fold while the growth rate of the fruitworm is severely depressed. In tomato fruit, the levels of PPO are highest in small immature fruit but are essentially negligible in mature fruit. The growth rate of larvae on fruit was also negatively correlated with PPO activity, with the fastest larval growth rate occurring when larvae fed on mature fruit. The reduction in larval growth is proposed to result from the alkylation of amino acids/protein byo-quinones, and the subsequent reduction in the nutritive quality of foliage. This alkylation reduces the digestibility of dietary protein and the bioavailability of amino acids. We believe that this mechanism of digestibility reduction may be extrapolatable to other plant-insect systems because of the ubiquitous cooccurrence of PPO and phenolic substrates among vascular plant species.     

多酚氧化酶催化单宁的氧化聚合反应研究

研究了多酚氧化酶催化单宁的氧化聚合反应情况 ,探索了反应条件及 7种化合物对酶活性的抑制效果 ,对产物进行了红外光谱和紫外光谱测试 ,相对粘度法测定了产物的分子量。结果表明 ,反应的最佳条件是pH为 5 .2 9,温度为 3 0℃ ,在所用抑制剂中 ,硫酸铜的抑制作用最强 ,与单宁酸的IR图相比 ,产物的 -OH吸收带变窄 ,在 12 60cm-1处吸收峰增强 ,而紫外吸收无变化。产物平均分子量为 3 5 41,说明多酚氧化酶能催化单宁的氧化聚合     

Effects of allelochemicals on plant respiration and oxygen isotope fractionation by the alternative oxidase

The goal of this investigation was to determine the effects of allelochemicals on plant respiration that thereby may be responsible for their role in growth inhibition. We have tested the effects of juglone, quercetin, cinnamic acid, and-pinene on respiration rates, and electron partitioning through the cytochrome and alternative respiratory pathways, by measuring on-line oxygen consumption and oxygen isotope fractionation in soybean cotyledon tissue. Cinnamic acid and-pinene decreased the oxygen consumption rate and increased the relative partitioning of electron transport to the alternative pathway. Possible biochemical mechanisms of these effects are discussed.     

过氧化氢在生成戊二酰-7-氨基头孢烷酸过程中的影响

头孢菌素C(CPC)在D-氨基酸氧化酶为催化剂的情况下,与O2反应生成α-ketoadipyl-7-ACA,再与过氧化氢反应生成戊二酰基-7-氨基头孢烷酸(Gl-7-ACA).在此反应中生成的α-ketoadipyl-7-ACA是造成目标产物收率低的关键因素之一.通过加入适量的双氧水可以使剩余的α-ketoadipl-7-ACA转化为Gl-7-ACA,从而使裂解的收率提高5%.     

Properties of cuticular oxidases used for sex pheromone biosynthesis byHeliothis zea

Biosynthesis of the aldehydic sex pheromone components released by females ofHeliothis zea was found to be catalyzed by primary alcohol oxidases residing in the cuticle that covers the glands. Activity, as indicated by conversion of primary alcohol to aldehyde, was as high in cell-free cuticle as it was in intact pheromone glands. Studies indicated that some activity was associated with the surface of the epicuticle and could be removed, into buffer, by sonication. However, the majority of activity lies within the inner epicuticle and exo- and endocuticular layers. The oxidase was not functional in pharate pupae that did not have mature adult cuticle but became functional just prior to adult emergence. The enzyme in individual glands was saturated at alcohol concentrations above 100 n. moles. Nonionic detergents did not affect the activity of the oxidase in the cuticle but treatment with either 7 M urea or 1% SDS resulted in total loss of activity. Studies on the effect of pH indicated an optimum at 6.4; however, activity was high throughout the range of 5–9. The oxidase was functional in both dichloromethane and hexane, suggesting that this enzyme system may have applications for organic synthesis of aldehydes.