Atomic-Level Regulation of Cobalt Single-Atom Nanozymes: Engineering High-Efficiency Catalase Mimics

Nanozymes aim to mimic the highly evolved active centers of natural enzymes. Despite progress in nanozyme engineering, their catalytic performance is much less favorable compared with natural enzymes. This study shows that precise control over the atomic configuration of the active centers of Co single-atom nanozymes (SAzymes) enables the rational regulation of their catalase-like performance guided by theorical calculations. The constructed Co-N₃PS SAzyme exhibits an excellent catalase-like activity and kinetics, exceeding the representative controls of Co-based SAzymes with different atomic configurations. Moreover, we developed an ordered structure-oriented coordination design strategy for rationally engineering SAzymes and established a correlation between the structure and enzyme-like performance. This work demonstrates that precise control over the active centers of SAzymes is an efficient strategy to mimic the highly evolved active sites of natural enzymes.     

Attachment of Nanoparticles to Pyrolytic Graphite Electrode and Its Application for the Direct Electrochemistry and Electrocatalytic Behavior of Catalase

Abstract

A highly hydrophilic, nontoxic, and conductive effect of colloidal gold nanoparticles (GNP) and multi-walled carbon nanotubes (MWCNT) on pyrolytic graphite electrode has been demonstrated. The direct electron transfer of catalase (CAT) was achieved based on the immobilization of MWCNT/CAT-GNP on a pyrolytic graphite electrode by a Nafion film. The immobilized catalase displayed a pair of well-defined and nearly reversible redox peaks in 0.1 M phosphate buffer solution (PBS) (pH 6.98). The dependence of E°′on solution pH indicated that the direct electron transfer reaction of catalase was a single-electron-transfer coupled with single-proton-transfer reaction process. The immobilized catalase maintained its biological activity, showing a surface controlled electrode process with an apparent heterogeneous electron transfer rate constant (k _s) of 1.387±0.1 s^?1 and charge-transfer coefficient (α) of 0.49, and displayed electrocatalytic activity in the electrocatalytic reduction of hydrogen peroxide. Therefore, the resulting modified electrode can be used as a biosensor for detecting hydrogen peroxide.     

新型兼具GPX,SOD, CAT活性的水杨醛Schiff碱衍生物的合成

以5-磺基水杨醛为母体, 经氯甲基化和硒化反应引入催化基团-SeH, 此化合物经空气氧化、Schiff碱反应以及锰螯合反应, 最终得到一种新型的水杨醛Schiff碱类谷胱甘肽过氧化物酶(GPX)模拟物. 测定了该模拟物的红外光谱、核磁共振谱、质谱和抗氧化能力. 此模拟物同时兼具超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性, 水溶性好, 保护线粒体免遭氧化损伤能力强, 具有作为药物前体的潜力.     

Detection of a novel familial catalase mutation (Hungarian type D) and the possible risk of inherited catalase deficiency for diabetes mellitus

The enzyme catalase is the main regulator of hydrogen peroxide metabolism. Recent findings suggest that a low concentration of hydrogen peroxide may act as a messenger in some signalling pathways whereas high concentrations are toxic for many cells and cell components. Acatalasemia is a genetically heterogeneous condition with a worldwide distribution. Yet only two Japanese and three Hungarian syndrome-causing mutations have been reported. A large-scale (23 130 subjects) catalase screening program in Hungary yielded 12 hypocatalasemic families. The V family with four hypocatalasemics (60.6 +/- 7.6 MU/L) and six normocatalasemic (103.6 +/- 23.5 MU/L) members was examined to define the mutation causing the syndrome. Mutation screening yielded four novel polymorphisms. Of these, three intron sequence variations, namely G-->A at the nucleotide 60 position in intron 1, T-->A at position 11 in intron 2, and G-->T at position 31 in intron 12, are unlikely to be responsible for the decreased blood catalase activity. However, the novel G-->A mutation in exon 9 changes the essential amino acid Arg 354 to Cys 354 and may indeed be responsible for the decreased catalase activity. This inherited catalase deficiency, by inducing an increased hydrogen peroxide steady-state concentration in vivo, may be involved in the early manifestation of type 2 diabetes mellitus for the 35-year old proband.     

硝基芳烃对鲤鱼肝脏过氧化氢酶的抑制及构效关系研究

在离体实验条件下，测定了１４种硝基芳烃化合物对鲤鱼肝脏过氧化氢酶的半抑制浓度ＥＣ５０值，根据－ｌｇＥＣ５０值，以一阶价分子连接性指数＾１Ｘ＾ｖ、取代基数总和Σσ＾－、指示变量Ｉ、分子分支指数＾１Ｋα、分子最低空轨道能ＥＬＵＭＯ及辛醇－水分配系数ｌｇＰ为参数，进行定量结构活性相关性研究。     

Amperometric Determination of Catalase in Brazilian Commercial Honeys

A sensor for catalase (CAT) amperometric detection in association with flow injection analysis and a tubular reactor containing Amberlite IRA-743 resin was developed. Catalase was quantified in 14 samples of Brazilian commercial honeys. At flow rate of 1.5 mL min^?1 and injecting 200 µL of sample volumes, a sampling frequency of 90 determinations per hour was afforded. The linear dynamic range in CAT extends from 50 to 5000 UI mL^?1, at pH 7.4. The levels of CAT vary from 9.97 to 99.07 UI mg^?1. Taking into account these results, an inverse correlation was obtained with CAT and H₂O₂ levels.     

Activation‐Based Catalase Enzyme Electrode and its Usage for Glucose Determination

Abstract

In this study, a novel type amperometric biosensor, which is based on the activation of catalase enzyme by glucose, was developed and used for the sensitive determination of glucose. For the preparation of the biosensor catalase enzyme was immobilized in gelatin by using cross‐linking agent glutaraldehyde and fixed on a pretreated teflon membrane of a dissolved oxygen probe. Glucose was used as an activator for the catalase enzyme and determination of glucose is based on the assay of the differences on the catalase activity of the biosensor on the oxygenmeter in the absence and the presence of glucose in the reaction medium. The responses of the activation based catalase biosensor have a linear relation to glucose concentrations and good measurement correlation between 0.5 and 5.0 µM with 2 min response time. In the optimization studies of the biosensor the most suitable catalase amount were found as 1324 U cm^?2 and also phosphate buffer (pH: 7.0; 50 mM) and 35°C were obtained as the optimum working conditions. For the characterization studies of the biosensor some parameters such as activator and interference effects of some substances on the biosensor response, reproducibility and operational stability were performed.     

荧光光谱法研究除草剂扑草净和扑灭通与过氧化氢酶的相互作用

应用荧光光谱法研究了水溶液中三嗪类除草剂扑草净和扑灭通与过氧化氢酶分子间的相互作用。结果表明,除草剂对过氧化氢酶的荧光均有较强的猝灭作用,且静态猝灭是引起CAT荧光猝灭的主要原因。从荧光猝灭结果求出除草剂和CAT的结合常数及结合位点数。扑灭通:K=6.17×10~6 L·mol-1,n=1.45;扑草净:K=2.12×10~5 L·mol-1,n=1.19。并依据能量转移机制,求出了扑灭通、扑草净和CAT相互结合时,其给体-受体间距离r分别为0.140和0.155 nm。由此可见,扑灭通与CAT的结合作用要强于扑草净,并推测出除草剂与CAT的Tyr214发生结合作用。     

偏最小二乘红外光谱法及其对三种过氧化氢酶二级结构的测定

以九种蛋白质及其晶体结构数据为校准，建立了蛋白质二级结构测定的偏最小二乘红外光谱定量法，并分析了三种过氧化氢酶在水溶液中的二级结构，结果表明本法对于测定水溶液中二级结构是一种简便，有效，准确和可靠的方法。     

污泥对土壤过氧化氢酶活性的影响

选取自来水厂污泥为实验样品,采用实验室模拟的方法研究了污泥对土壤过氧化氢酶活性及热力学特征参数的影响。结果表明：在不同污泥含量下所有测定期间内过氧化氢酶活性都呈现抑制作用,污泥含量越高,对过氧化氢酶的抑制作用越明显。含污泥的土壤从第1d到第7d,土壤酶活性迅速降低,表现出明显的抑制作用;在第7d,污泥对过氧化氢酶活性的抑制作用达到最大,之后过氧化氢酶的活性缓慢恢复,28d后酶活性趋于平缓。热力学研究表明,过氧化氢酶最适宜温度为307—327K。土壤中含污泥时所需的Ea和ΔH比CK高,且污泥含量越大所需的Ea和ΔH越高,说明酶促反应越来越困难。