Laccase-based amperometric biosensor enhanced by Ni/Au/PPy-COOH multisegmental nanowires for selective dopamine detection

One-dimensional Ni/Au/PPy-COOH nanowires with multiple segments were synthesized in this study. Smooth surfaces and magnetic properties of nanowires were investigated by scanning transmission electron microscopy (SEM), Energy-dispersive X-ray spectroscopy (EDX), and Electron Spin Resonance (ESR) techniques. The nanowires were used to modify the screen-printed electrode surface and as a micro-environment for Trametes versicolor laccase. The ability of this enzyme biosensor to detect dopamine change in human biological samples was demonstrated by a wide linear range (0.01–50 μM) and a low LOD (2.265 nM). In addition, the biosensor exhibited excellent selectivity allowing the detection of dopamine in the presence of ascorbic acid, uric acid, L-Cys, serotonin, and glucose, with high sensitivity of reduction currents obtained at −0.2 V (vs. Ag/AgCl). The proposed biosensor allowed the detection of dopamine in commercial serum and artificial urine with recovery values close to 100 %. It also demonstrated reproducibility, reusability, and long-term storage stability. The sensitivity, K_m^app, and I_max values of the biosensor were determined as 2.05 μM and 1.03 μA, respectively. The LAC-Ni/Au/PPy-COOH/NAF/SPE biosensor is a reliable design for detecting dopamine with a wide linear range.     

两种功能化纳米金粒子固酶电极的催化性能

以合成的4-巯基苯甲酸功能化纳米金粒子和聚乙烯基吡啶包覆纳米金粒子分别作为固酶载体,制备了2种新型固酶电极,在此基础上组装了2种酶燃料电池。采用电化学方法结合紫外可见分光光度法、透射电镜技术等手段研究了固酶载体的形貌,酶-载体间相互作用对电极表面固定酶分子的光谱学性质,酶-电极间直接电子迁移能力和催化底物反应性能的影响,进一步评估和比较了两种酶燃料电池的能量输出性能。实验结果表明：4-巯基苯甲酸功能化纳米金粒子固酶基电极可以实现酶-电极间的直接电子迁移而且对葡萄糖和氧气具有良好的催化性能(催化反应起始电位分别为-0.03和0.96 V,底物转化频率分别是1.3和0.5 s^-1),其催化性能的重现性、长期使用性能、酸碱耐受性和热稳定性良好,随着自组装固酶层数的增加,催化性能随之增强直至达到极限催化电流;电池性能测试结果表明4-巯基苯甲酸功能化纳米金粒子固酶基燃料电池的开路电压为0.88 V,最大输出能量密度：864.0 μW·cm^-2,长期使用性能优异(储存3 周后仍可达到最佳能量输出的80%以上)。     

Surface hydrophobization of cotton via laccase-mediated polydopamine deposition and dodecyl gallate grafting

Natural fibers containing components with phenolic hydroxyl groups, such as jute, wool, and silk, can be directly modified by laccase-catalyzed grafting. However, cellulosic fibers like cotton cannot be functionalized in this manner. In this work, we developed a facile two-step method to graft polymers on cotton fabric via laccase catalysis. First, polydopamine (PDA) coating was deposited on the surface of the cotton fabrics via catalysis of laccase/2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) system. Then, the newly formed PDA coating acted as the secondary reaction platform for subsequent laccase-mediated grafting of hydrophobic monomer dodecyl gallate (DG). The oxidation of dopamine (DA) catalyzed with the laccase/TEMPO system was investigated using UV–visible (UV–vis) spectroscopy. The scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) results verified that the PDA was coated on the surface of cotton fibers. Fourier transform infrared (FTIR) spectra indicated that the PDA-coated cotton was successfully grafted with DG (DG-PDA-cotton). According to the weighting method, the grafting percentage was about 1.06%. The hydrophobicity of the DG-PDA-cotton fabrics was greatly improved with a contact angle of 133°. Also, the grafted cotton fabrics show repellency of water-soluble stains like coffee, milk, and tea. This study provides a new strategy for surface modification of cotton by laccase-mediated grafting, which offers the references for the green fabrication of cotton fabrics with improved functionalization.     

Syntheses of aromatic aldehydes by laccase without the help of mediators

This communication reports the selective bioconversions of substituted toluenes to substituted benzaldehydes without the help of any mediators by purified laccase of indigenous fungal strain Fomes durissimus microbial type culture collection (MTCC)-1173. Molecular mass of laccase purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was found to be 74.86 kDa (～75 kDa). By using this purified laccase, selective bioconversions of 3-nitrotoluene to 3-nitrobenzaldehyde, 2-fluorotoluene to 2-fluorobenzaldehyde, 4-fluorotoluene to 4-fluorobenzaldehyde, 2-chlorotoluene to 2-chlorobenzaldehyde and 4-chlorotoluene to 4-chlorobenzaldehyde have been done without the help of mediator molecules within 1–2 hrs at room temperature and pressure with high yields (>90%). All the above bioconversions are good examples of green chemistry.     

L-半胱氨酸/漆酶修饰的纳米金阴极对氧还原的电催化

以四氯金酸和L-半胱氨酸为原料,合成了L-半胱氨酸功能化纳米金粒子,将此纳米粒子修饰在金盘电极表面并共价偶联漆酶分子。 以循环伏安法研究了此固定漆酶电极在无氧磷酸盐缓冲液中电化学行为和催化氧还原能力,并进一步评估了其作为氧传感器使用的性能：以计时电流法测定其对氧气的检测限,与氧气的亲和力（以米氏常数表征）,研究了传感器的长期使用性、热稳定性和pH-催化电流关系。 结果表明,此固定漆酶电极可以实现漆酶活性中心T2与导电纳米粒子间的直接电子迁移而无需任何外加电子中介体（氧化还原峰的式电位为192.5 mV vs AgCl/Ag）,并在接近漆酶活性中心T3氧化还原式电位（780 mV vs NHE）附近发生氧还原;测得的固酶电极与氧气的亲和力较高（米氏常数为216.4 μmol/L）且检测限低达0.22 μmol/L,在4 ℃下保存60 d后活力仍然保持初始活力的大约78%。 但这种电极的热稳定性较差,受pH值影响较明显,在pH值近于生理条件时几乎完全丧失活力。     

Laccase Encapsulation in ZIF-8 Metal-Organic Framework Shows Stability Enhancement and Substrate Selectivity

CgL1 laccase from Corynebacterium glutamicum was encapsulated into the metal-organic framework (MOF) ZIF-8 which was synthesized in a rapid enzyme friendly aqueous synthesis, the fastest in situ encapsulation of laccases reported to date. The obtained enzyme/MOF, i. e. laccase@ZIF-8 composite showed enhanced thermal (up to 70 °C) and chemical (N,N-dimethylformamide) stability, resulting in a stable heterogenous catalyst, suitable for high temperature reactions in organic solvents. Furthermore, the defined structure of ZIF-8 produced a size selective substrate specificity, so that substrates larger than the pore size were not accepted. Thereby, 2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) was used to verify that the enzyme is immobilized inside the MOF versus the outside surface. The enzyme@MOF composite was analyzed by atomic absorption spectroscopy (ASS) to precisely determine the enzyme loading to 2.1 wt%.     

反相胶束介质中漆酶催化反应中间体信息的捕捉及其解析

利用反相胶束独特的介质特性，捕捉到漆酶催化反应的中间体光谱动力学信号，实现了对酶催化反应过程的在线光谱研究．通过解析反应过程中在线测得的动力学光谱数据矩阵，解得中间体的生成和衰减均为一级反应，并求出实验条件下的反应速率常数．通过实验数据与解析结果的比较证实了方法的合理性．     

磁性壳聚糖微球的制备及其用作漆酶固定化载体

磁性壳聚糖微球的制备及其用作漆酶固定化载体;壳聚糖;固定化;漆酶;酶活力     

Laccase‐catalyzed polymerization of lignocatechol and affinity on proteins of resulting polymers

For the synthesis of a new biologically functional polymer from a natural resource by an environment‐friendly method, the laccase‐catalyzed polymerization of a lignin‐based macromonomer, lignocatechol, was carried out for the first time in ethanol–phosphate buffer solvent system to give crosslinked polymers in good yields. Lignocatechol was prepared by the phase separation system of lignin and catechol in aqueous sulfuric acid. The copolymerization was also performed with urushiol to afford the corresponding copolymers in high yields. The polymerization mechanism was estimated by the IR and pyrolysis GC‐MS measurements, suggesting that the polymerization proceeded mainly at the catechol ring through a quinone radical intermediate. The thermal properties were measured by the DSC, TG, and TMA analyses, indicating that the polymers had high thermal stabilities because of the crosslinked structures. In addition, it was found that the resulting polymers had the affinity of bovine serum albumin (BSA) and glucoamylase. © 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 824–832, 2009