Masking the Peroxidase-Like Activity of the Molybdenum Disulfide Nanozyme Enables Label-Free Lipase Detection

Two-dimensional MoS₂ nanoparticles (2D-nps) exhibit artificial enzyme properties that can be regulated at bio-nanointerfaces. We discovered that protein lipase is able to tune the peroxidase-like activity of MoS₂ 2D-nps, offering low-nanomolar, label-free detection and identification in samples with unknown identity. The inhibition of the peroxidase-like activity of the MoS₂ 2D-nps was demonstrated to be concentration dependent, and as low as 5 nm lipase was detected with this approach. The results were compared with those obtained with several other proteins that did not display any significant interference with the nanozyme behavior of the MoS₂ 2D-nps. This unique response of lipase was characterized and exploited for the successful identification of lipase in six unknown samples by using qualitative visual inspection and a quantitative statistical analysis method. The developed methodology in this approach is noteworthy for many aspects; MoS₂ 2D-nps are neither labeled with a signaling moiety nor modified with any ligands for signal readout. Only the intrinsic nanozyme activity of the MoS₂ 2D-nps is exploited for this detection approach. No analytical equipment is necessary for the visual detection of lipase. The synthesis of the water-soluble MoS₂ 2D-nps is low costing and can be performed in bulk scale. Exploring the properties of 2D-nps and their interactions with biological materials reveals highly interesting yet instrumental features that offer the development of novel bioanalytical approaches.     

Effect of short-term storage on phenolic content,o-diphenolase and peroxidase activities of cut yam tubers (Dioscorea sp)

The effect of short-term storage on the protein, phosphorus and phenolic content as well as peroxidase and o-diphenolase activities of cut, harvested Jamaican yam (Dioscorea sp) tubers (D rotundata. D alata and D cayenensis) was studied. There was an initial increase in the total phenolic content up to the third week of storage followed by a gradual decrease to the sixth week. Phenolic content was found to be highest in D cayenensis followed by D rotundata and D alata. The activities of peroxidase (EC 1. 11. 1. 7) and o-diphenolase (EC 1. 10.3.1) increased steadily up to the third week of storage and thereafter decreased to the fifth week. The intensity and rapidity of browning in tubers when cut, correlated very closely with the tuber o-diphenolase and phenolic content levels while the onset of rotting correlated with the peroxidase activity levels in the species studied.     

Effect of surfactants on phenol removal by the method of polymerization and precipitation catalysed by Coprinus cinereus peroxidase

The removal of phenol by peroxidase‐catalysed polymerization was examined using Coprinus cinereus peroxidase in the presence of surfactants. The non‐ionic surfactants with poly(oxyethene) residues, Triton X‐100, Triton X‐405 and Tween 20, enhanced the phenol removal efficiency at a level similar to high relative molecular mass poly(ethylene glycol) (relative molecular mass 3000). Although the improvement in the removal efficiency was less than that of Triton X‐100, Span 20, sodium lauryl sulfate (SDS) and lauryl trimethylammonium bromide (DTAB) also enhanced the removal efficiency. The requirement of the enzyme for almost 100% removal of 100 mg dm^?3 phenol decreased to one‐fourth by the addition of 30 mg dm^?3 Triton X‐100. Triton X‐100, Triton X‐405, Tween 20 and DTAB could reactivate the enzyme precipitated with the phenol polymer, leading to the restarting of the phenol removal reaction. Copyright © 2003 Society of Chemical Industry     

Induction of favism-like symptoms in the rat: Effects of vicine and divicine in normal and buthionine sulphoximine-treated rats

Rats were fed an adequate or a deficient diet and offered water or buthionine sulphoximine (BSO) solution for 2 weeks, and then the same diets with vicine for another week in experiment 1. BSO in combination with the deficient diet caused a marked decrease in blood glutathione (GSH) and growth retardation but failed to show any effects resulting from supplementation with vicine. In experiment 2 the rats were given an adequate diet and BSO as before, and injected intravenously with divicine (DV). Here again, BSO depressed rat growth, and so did DV. Each of the insults also caused haematological changes, especially a fall in GSH, but the most severe changes appeared in the group treated with both BSO and DV. A decrease in haematocrit and increases in adrenal and spleen weight were also noted. In experiment 3 the rats were injected with different doses of DV, without pretreatment with BSO. The main effect was a drop in blood GSH and haematocrit, and an increase in adrenal and spleen weights, all of which were dose-related. Administration of the higher doses of DV resulted in a severe cyanosis followed by death within a relatively short period of time.     

Effect of H2O2 addition mode on enzymatic removal of phenol from wastewater in the presence of polyethylene glycol

The effect of batch and semi-batch additions of the stoichiometric amount of H₂O₂ on enzymatic removal of phenol has been studied over the phenol concentration range of 1-10 mmol/L. It is found that the ratio between the maximum H₂O₂ concentration during the reaction and the initial horseradish peroxidase concentration ((H₂O₂)_max/(HRP)_o) controls the rate of phenol removal by horseradish peroxidase in the presence of the additive, polyethylene glycol. An optimum range of this ratio is determined to be between 10 and 25 μmol/U. Empirical models are proposed to predict the values of (H₂O₂)_max/(HRP)_o required to treat given phenol concentrations under different modes for adding H₂O₂.     

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.     

辣根过氧化物酶在有机合成中的应用

综述了辣根过氧化物酶在有机合成中作为催化剂使用,其催化机理为过氧化循环过程;以及辣根过氧化物酶催化合成苯胺、苯胺衍生物、酚类的研究进展,其特点为催化效率高。用酶的催化聚合作用来处理酚类、芳香胺类化合物的污染,显示了广阔的应用前景。     

基于金纳米-植酸胶束混合组装的过氧化氢传感器的电化学研究

采用混合组装技术,利用植酸胶束(IP6micelles)的磷酸酯键络合辣根过氧化物酶(HRP)和金纳米粒子(GNPs),形成了具有生物亲和性的纳米复合材料,保持了辣根过氧化物酶的生物活性,并利用金纳米粒子的高电子密度、介电特性和催化性能,实现了HRP与玻碳电极(GCE)表面的直接电子转移。Nafion膜的滴加能提高电极的选择性和稳定性。实验过程中借助紫外-可见吸收光谱和透射电子显微镜进行表征,实验结果证明:GNPs的高导电和高催化性能,结合植酸胶束的优良生物相容性和对酶的高负载量的特点,使得吸附在其上的HRP保持活性,制备的生物传感器能对H2O2进行电催化还原。Nafion/HRP-IP6micelles-GNPs/GCE对H2O2的线性浓度范围为5×10-7～1.15×10-5mol/L(线性相关系数r=0.993,n=9),最低检测限为0.1μmol/L(信噪比S/N=3),米氏常数为0.002 4 mmol/L。